Classifications

• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B04—CENTRIFUGAL APPARATUS OR MACHINES FOR CARRYING-OUT PHYSICAL OR CHEMICAL PROCESSES
  • B04B—CENTRIFUGES
  • B04B15/00—Other accessories for centrifuges
  • B04B15/02—Other accessories for centrifuges for cooling, heating, or heat insulating
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
  • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
  • A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
  • A61M1/3616—Batch-type treatment
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
  • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
  • A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
  • A61M1/3693—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits using separation based on different densities of components, e.g. centrifuging
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
  • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
  • A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
  • A61M1/3693—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits using separation based on different densities of components, e.g. centrifuging
  • A61M1/3696—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits using separation based on different densities of components, e.g. centrifuging with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B04—CENTRIFUGAL APPARATUS OR MACHINES FOR CARRYING-OUT PHYSICAL OR CHEMICAL PROCESSES
  • B04B—CENTRIFUGES
  • B04B11/00—Feeding, charging, or discharging bowls
  • B04B11/04—Periodical feeding or discharging; Control arrangements therefor
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B04—CENTRIFUGAL APPARATUS OR MACHINES FOR CARRYING-OUT PHYSICAL OR CHEMICAL PROCESSES
  • B04B—CENTRIFUGES
  • B04B13/00—Control arrangements specially designed for centrifuges; Programme control of centrifuges
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B04—CENTRIFUGAL APPARATUS OR MACHINES FOR CARRYING-OUT PHYSICAL OR CHEMICAL PROCESSES
  • B04B—CENTRIFUGES
  • B04B5/00—Other centrifuges
  • B04B5/04—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers
  • B04B5/0407—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers for liquids contained in receptacles
  • B04B5/0428—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers for liquids contained in receptacles with flexible receptacles
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B04—CENTRIFUGAL APPARATUS OR MACHINES FOR CARRYING-OUT PHYSICAL OR CHEMICAL PROCESSES
  • B04B—CENTRIFUGES
  • B04B5/00—Other centrifuges
  • B04B5/04—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers
  • B04B5/0442—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B04—CENTRIFUGAL APPARATUS OR MACHINES FOR CARRYING-OUT PHYSICAL OR CHEMICAL PROCESSES
  • B04B—CENTRIFUGES
  • B04B9/00—Drives specially designed for centrifuges; Arrangement or disposition of transmission gearing; Suspending or balancing rotary bowls
  • B04B9/08—Arrangement or disposition of transmission gearing ; Couplings; Brakes
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B04—CENTRIFUGAL APPARATUS OR MACHINES FOR CARRYING-OUT PHYSICAL OR CHEMICAL PROCESSES
  • B04B—CENTRIFUGES
  • B04B9/00—Drives specially designed for centrifuges; Arrangement or disposition of transmission gearing; Suspending or balancing rotary bowls
  • B04B9/10—Control of the drive; Speed regulating
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
  • A61M2202/00—Special media to be introduced, removed or treated
  • A61M2202/04—Liquids
  • A61M2202/0413—Blood
  • A61M2202/0427—Platelets; Thrombocytes
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
  • A61M2205/00—General characteristics of the apparatus
  • A61M2205/33—Controlling, regulating or measuring
  • A61M2205/3306—Optical measuring means
  • A61M2205/331—Optical measuring means used as turbidity change detectors, e.g. for priming-blood or plasma-hemoglubine-interface detection
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
  • A61M2205/00—General characteristics of the apparatus
  • A61M2205/33—Controlling, regulating or measuring
  • A61M2205/3368—Temperature
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B04—CENTRIFUGAL APPARATUS OR MACHINES FOR CARRYING-OUT PHYSICAL OR CHEMICAL PROCESSES
  • B04B—CENTRIFUGES
  • B04B5/00—Other centrifuges
  • B04B5/04—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers
  • B04B5/0442—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation
  • B04B2005/045—Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation having annular separation channels
• • B—PERFORMING OPERATIONS; TRANSPORTING
  • B04—CENTRIFUGAL APPARATUS OR MACHINES FOR CARRYING-OUT PHYSICAL OR CHEMICAL PROCESSES
  • B04B—CENTRIFUGES
  • B04B13/00—Control arrangements specially designed for centrifuges; Programme control of centrifuges
  • B04B2013/006—Interface detection or monitoring of separated components

Definitions

• This invention relates to novel methods, devices and apparatuses for the centrifugal separation of a liquid into its components of varying specific gravities, and is more particularly directed toward a blood separation device useful, for example, in the separation of blood components for use in various therapeutic regimens.
• Centrifugation utilizes the principle that particles suspended in solution will assume a particular radial position within the centrifuge rotor based upon their respective densities and will therefore separate when the centrifuge is rotated at an appropriate angular velocity for an appropriate period of time.
• Centrifugal liquid processing systems have found applications in a wide variety of fields. For example, centrifugation is widely used in blood separation techniques to separate blood into its component parts, that is, red blood cells, platelets, white blood cells, and plasma.
• the liquid portion ofthe blood is a protein-salt solution in which red and white blood cells and platelets are suspended.
• Plasma which is 90 percent water, constitutes about 55 percent of the total blood volume.
• Plasma contains albumin (the chief protein constituent), fibrinogen (responsible, in part, for the clotting of blood), globulins (including antibodies) and other clotting proteins.
• Plasma serves a variety of functions, from maintaining a satisfactory blood pressure and providing volume to supplying critical proteins for blood clotting and immunity. Plasma is obtained by separating the liquid portion of blood from the cells suspended therein.
• Red blood cells erythrocytes
• Red blood cells contain hemoglobin, a complex iron-containing protein that carries oxygen throughout the body while giving blood its red color. The percentage of blood volume composed of red blood cells is called the "hematocrit.”
• White blood cells are responsible for protecting the body from invasion by foreign substances such as bacteria, fungi and viruses.
• white blood cells exist for this purpose, such as granulocytes and macrophages which protect against infection by surrounding and destroying invading bacteria and viruses, and lymphocytes which aid in the immune defense.
• Platelets are very small cellular components of blood that help the clotting process by sticking to the lining of blood vessels. Platelets are vital to life, because they help prevent both massive blood loss resulting from trauma and blood vessel leakage that would otherwise occur in the course of normal, day-to-day activity.
• whole blood is collected and prevented from clotting by the addition of an appropriate anticoagulant, it can be centrifuged into its component parts. Centrifugation will result in the red blood cells, which weigh the most, packing to the most outer portion ofthe rotating container, while plasma, being the least dense will settle in the central portion of the rotating container. Separating the plasma and red blood cells is a thin white or grayish layer called the buffy coat.
• the buffy coat layer consists ofthe white blood cells and platelets, which together make up about 1 percent ofthe total blood volume.
• red blood cells are routinely transfused into patients with chronic anemia resulting from disorders such as kidney failure, malignancies, or gastrointestinal bleeding and those with acute blood loss resulting from trauma or surgery.
• the plasma component is typically frozen by cryoprecipitation and then slowly thawed to produce cryoprecipitated antihemophiliac factor (AHF) which is rich in certain clotting factors, including Factor VIII, fibrinogen, von Willebrand factor and Factor XIII.
• AHF antihemophiliac factor
• Cryoprecipitated AHF is used to prevent or control bleeding in individuals with hemophilia and von Willebrand's disease.
• Platelets and white blood cells which are found in the buffy layer component, can be used to treat patients with abnormal platelet function (thrombocytopenia) and patients that are unresponsive to antibiotic therapy, respectively.
• Centrifugal systems also referred to as blood-processing systems
• discontinuous-flow systems whole blood from the donor or patient flows through a conduit into the rotor or bowl where component separation takes place.
• These systems employ a bowl-type rotor with a relatively large (typically 200 ml or more) volume that must be filled with blood before any ofthe desired components can be harvested.
• the bowl is full, the drawing of fresh blood is stopped, the whole blood is separated into its components by centrifugation, and the unwanted components are returned to the donor or patient through the same conduit intermittently, in batches, rather than on a continuous basis.
• whole blood is again drawn from the donor or patient, and a second cycle begins. This process continues until the required amount ofthe desired component has been collected.
• Discontinuous-flow systems have the advantage that the rotors are relatively small in diameter but have the disadvantage that the extracorporeal volume (i.e., the amount of blood that is out ofthe donor at any given time during the process) is large. This, in turn, makes it difficult or impossible to use discontinuous systems on people whose size and weight will not permit the drawing ofthe amount of blood required to fill the rotor.
• Discontinuous-flow devices are used for the collection of platelets and/or plasma, and for the concentration and washing of red blood cells. They are used to reconstitute previously frozen red blood cells and to salvage red blood cells lost intraoperatively.
• a discontinuous-flow system is disclosed by McMannis, et al., in his U.S. Patent No. 5,316,540, and is a variable volume centrifuge for separating components of a fluid medium, comprising a centrifuge that is divided into upper and lower chambers by a flexible membrane, and a flexible processing container bag positioned in the upper chamber ofthe centrifuge.
• the McMannis, et al system varies the volume ofthe upper chamber by pumping a hydraulic fluid into the lower chamber, which in turn raises the membrane and squeezes the desired component out ofthe centrifuge.
• the McMannis, et al, system takes up a fairly large amount of space, and its flexible pancake-shaped rotor is awkward to handle.
• the McMannis, et al, system does not permit the fluid medium to flow into and out of the processing bag at the same time, nor does it permit fluid medium to be pulled out ofthe processing bag by suction.
• whole blood from the donor or patient also flows through one conduit into the spinning rotor where the components are separated.
• the component of interest is collected and the unwanted components are returned to the donor through a second conduit on a continuous basis as more whole blood is being drawn. Because the rate of drawing and the rate of return are substantially the same, the extracorporeal volume, or the amount of blood that is out of the donor or patient at any given time in the procedure, is relatively small.
• These systems typically employ a belt- type rotor, which has a relatively large diameter but a relatively small (typically 100 ml or less) processing volume.
• continuous-flow systems have the advantage that the amount of blood that must be outside the donor or patient can be relatively small, they have the disadvantage that the diameter ofthe rotor is large. These systems are, as a consequence, large. Furthermore, they are complicated to set up and use. These devices are used almost exclusively for the collection of platelets.
• Continuous-flow systems are comprised of rotatable and stationary parts that are in fluid communication. Consequently, continuous-flow systems utilize either rotary seals or a J-loop.
• rotary centrifuge seals A variety of types of rotary centrifuge seals have been developed. Some examples of rotary centrifuge seals which have proven to be successful are described in U.S. Pat. Nos. 3,409,203 and 3,565,330, issued to Latham. In these patents, rotary seals are disclosed which are formed from a stationary rigid low friction member in contact with a moving rigid member to create a dynamic seal, and an elastomeric member which provides a resilient static seal as well as a modest closing force between the surfaces of the dynamic seal.
• One flow system heretofore contemplated to overcome the problem of the rotating seal utilizes a rotating carriage on which a single housing is rotatably mounted.
• An umbilical cable extending to the housing from a stationary point imparts planetary motion to the housing and thus prevents the cable from twisting.
• a family of dual member centrifuges can be used to effect cell separation.
• This type of centrifuge is disclosed in U.S. Pat. No. RE 29,738 to Adams entitled "Apparatus for Providing Energy Communication Between a Moving and a Stationary Terminal".
• the Adams patent discloses a centrifuge having an outer rotatable member and an inner rotatable member.
• the inner member is positioned within and rotatably supported by the outer member.
• the outer member rotates at one rotational velocity, usually called “one omega,” and the inner rotatable member rotates at twice the rotational velocity ofthe outer housing or "two omega.” There is thus a one omega difference in rotational speed of the two members.
• the term “dual member centrifuge” shall refer to centrifuges ofthe Adams type.
• the dual member centrifuge ofthe Adams patent is particularly advantageous in that, as noted above, no seals are needed between the container of fluid being rotated and the non-moving component collection containers.
• the system of the Adams patent provides a way to process blood into components in a single, sealed, sterile system wherein whole blood from a donor can be infused into the centrifuge while the two members of the centrifuge are being rotated.
• the outer member ofthe Lolachi centrifuge is rotated by a single electric motor which is coupled to the internal rotatable housing by belts and shafts.
• U.S. Pat. No. 4,108,353 to Brown entitled “Centrifugal Apparatus With Oppositely Positioned Rotational Support Means” discloses a centrifuge structure of the Adams type which includes two separate electrical motors. One electric motor is coupled by a belt to the outer member and rotates the outer member at a desired nominal rotational velocity. The second motor is carried within the rotating exterior member and rotates the inner member at the desired higher velocity, twice that ofthe exterior member.
• U.S. Pat. No. 4,109,855 to Brown, et al, entitled “Drive System For Centrifugal Processing Apparatus” discloses yet another drive system.
• the system of the Brown, et al. patent has an outer shaft, affixed to the outer member for rotating the outer member at a selected velocity.
• An inner shaft, coaxial with the outer shaft, is coupled to the inner member.
• the inner shaft rotates the inner member at twice the rotational velocity as the outer member.
• a similar system is disclosed in U.S. Pat. No. 4,109,854 to Brown entitled “Centrifugal Apparatus With Outer Enclosure”.
• Whole blood that is to be separated into its components is commonly collected into a flexible plastic donor bag, and the blood is centrifuged to separate it into its components through a batch process. This is done by spinning the blood bag for a period of about 10 minutes in a large refrigerated centrifuge.
• the main blood constituents i.e., red blood cells, platelets and white cells, and plasma, having sedimented and formed distinct layers, are then expressed sequentially by a manual extractor in multiple satellite bags attached to the primary bag.
• haemapheresis technology in whole blood donation. This technique consists of drawing and extracting on-line one or more blood components while a donation is performed, and returning the remaining constituents to the donor.
• haemapheresis systems preclude their use by transfusion centers for routine whole blood collection.
• U.S. Pat. No. 5,316,540 to McMannis, et al discloses a centrifugal processing apparatus, wherein the processing chamber is a flexible processing bag which can be deformed to fill it with biological fluid or empty it by means of a membrane which forms part ofthe drive unit.
• the bag comprises a single inlet/outlet tubing for the introduction and removal of fluids to the bag, and consequently cannot be used in a continual, on-line process.
• the processing bag has a the disadvantage of having 650 milliliter capacity, which makes the McMannis, et al, device difficult to use as a blood processing device.
• a bioadhesive sealant also referred to as a fibrin glue
• fibrin glue is a relatively new technological advance which attempts to duplicate the biological process ofthe final stage of blood coagulation.
• Clinical reports document the utility of fibrin glue in a variety of surgical fields, such as, cardiovascular, thoracic, transplantation, head and neck, oral, gastrointestinal, orthopedic, neurosurgical, and plastic surgery. At the time of surgery, the two primary components comprising the fibrin glue, fibrinogen and thrombin, are mixed together to form a clot.
• fibrin glue has its ability to: (1) achieve haemostasis at vascular anastomoses particularly in areas which are difficult to approach with sutures or where suture placement presents excessive risk; (2) control bleeding from needle holes or arterial tears which cannot be controlled by suturing alone; and (3) obtain haemostasis in heparinized patients or those with coagulopathy. See, Borst, H.G., et al, J Thorac. Cardiovasc. Surg., 84:548-553 (1982); Walterbusch, G. J, et al, Thorac. Cardiovasc. Surg., 30:234-23 5 (1982); and Wolner, F. J, et al, Thorac. Cardiovasc. Surg., 30:236-237 (1982).
• lipid-enveloped viruses such as HIV, associated with AIDS, cytomegalovirus (CMV), as well as Epstein-Barr virus and the herpes simplex viruses in fibrinogen preparations makes it unlikely that there will be a change in this policy in the foreseeable future.
• CMV cytomegalovirus
• human thrombin is also not currently authorized for human use in the United States. Bovine thrombin, which is licensed for human use in the United States is obtained from bovine sources which do not appear to carry significant risks for HIV and hepatitis, although other bovine pathogens, such as bovine spongiform and encephalitis, may be present. There have been a variety of methods developed for preparing fibrin glue.
• Rose, et al in U.S. Pat. No. 4,627,879 discloses a method of preparing a cryoprecipitated suspension containing fibrinogen and Factor XIII useful as a precursor in the preparation of a fibrin glue which involves (a) freezing fresh frozen plasma from a single donor such as a human or other animal, e.g. a cow, sheep or pig, which has been screened for blood transmitted diseases, e.g. one or more of syphilis, hepatitis or acquired immune deficiency syndrome, at about 80° C for at least about 6 hours, preferably for at least about 12 hours; (b) raising the temperature ofthe frozen plasma, e.g.
• the fibrin glue is then prepared by applying a defined volume ofthe cyroprecipitate suspension described above and applying a composition containing a sufficient amount of thrombin, e.g. human, bovine, ovine or porcine thrombin, to the site so as to cause the fibrinogen in the suspension to be converted to the fibrin glue which then solidifies in the form of a gel.
• thrombin e.g. human, bovine, ovine or porcine thrombin
• a second technique for preparing fibrin glue is disclosed by Marx in his U.S. Patent Number 5,607,694. Essentially, a cryoprecipitate as discussed previously serves as the source of the fibrinogen component and then Marx adds thrombm and liposomes.
• a fibrin glue prepared by mixing bovine thrombin not only with human coagulant proteins, such as fibrinogen, fibronectin, Factor XIII, and plasminogen, but also with bovine aprotinin and calcium chloride.
• fibrin glue is comprised primarily of fibrinogen and thrombin thus lacking an appreciable quantity of platelets. Platelets contain growth factors and healing factors which are assumed to be more prevalent in a platelet concentrate. Moreover, platelets aid in acceleration ofthe clotting process.
• centrifugal cell processing system wherein multiple batches of cells can be simultaneously and efficiently processed without the use of rotational coupling elements.
• a platelet concentrate that aids in increasing the rate of fibrin clot formation, thereby facilitating haemostasis.
• the apparatus will be essentially self-contained.
• the equipment needed to practice the method will be relatively inexpensive and the blood contacting set will be disposable each time the whole blood has been separated.
• one object of this invention is to provide a method and apparatus for the separation of components suspended or dissolved in a fluid medium by centrifugation. More specifically, one object of this invention is to provide a method for the separation and isolation of one or more whole blood components, such as platelet rich plasma, white blood cells and platelet poor plasma, from anticoagulated whole blood by centrifugation, wherein the components are isolated while the centrifuge is rotating.
• whole blood components such as platelet rich plasma, white blood cells and platelet poor plasma
• Another object of this invention is to utilize the isolated cell components in a therapeutic regimen.
• Another object of this invention is to provide an apparatus for the separation of whole blood components, wherein the apparatus contains a centrifuge bag that provides for simultaneous addition of whole blood from a source container and the withdrawal of a specific blood component during centrifugation.
• Another object of this invention is to provide disposable, single-use centrifuge bags for holding whole blood during the separation of components ofthe whole blood by centrifugation, wherein the bag is adapted for use in a portable, point-of-use centrifuge.
• one embodiment of this invention comprises a flexible, disposable centrifuge bag adapted to be rotated about an axis, comprising: a) one or more tubes, and b) upper and lower flexible sheets, each sheet having a doughnut shaped configuration, an inner perimeter defining a central core and an outer perimeter, wherein the upper and lower sheets are superimposed and completely sealed together at their outer perimeters, and wherein the tubes are sandwiched between the upper and lower sheets and extend from the central core toward the outer perimeter, such that when the upper and lower sheets are sealed at the inner perimeter the tubes are sealed between the upper and lower sheets at the inner perimeter and are in fluid communication with the environment inside and outside the centrifuge
• another embodiment ofthe present invention comprises a rigid molded container adapted to be rotated about an axis, comprising a rigid, annular body having an axial core that is closed at the top end and opened at the bottom end.
• the rigid molded container further comprises an interior collection chamber for receiving and holding a fluid medium to be centrifuged, the chamber having an outer perimeter, an inner perimeter, and a generally off-centered "figure eight" shaped cross-sectional area.
• the rigid molded container further comprises a first channel which extends radially from the core and is in fluid communication with a point near the outer perimeter ofthe chamber, and a second channel which extends radially from the core and is in fluid communication with an area near the narrow portion or "neck" ofthe figure eight-shaped chamber.
• the first and second channels thus provide fluid communication with the environment inside and outside the interior collection chamber.
• the first and second channels are fluidly connected to a dual lumen tubing having an inlet lumen and an outlet lumen.
• another embodiment ofthe present invention is an apparatus and method for separating components contained in a fluid medium. More particularly, the present invention utilizes the principles of centrifugation to allow for the separation of whole blood into fractions such as platelet rich plasma and platelet poor plasma.
• the above-described separation ofthe components is provided by utilizing a rotatable centrifuge motor comprising a base having a central column and a disposable centrifuge bag having a central core and which is positionable within the centrifuge motor and rotatable therewith.
• the disposable centrifuge bag which holds the whole blood during centrifugation, further comprises an inlet tube for introducing the whole blood to the centrifuge bag, and an outlet tube for removing the desired blood fraction from the centrifuge bag.
• the inlet and outlet tubes are in fluid communication with a dual lumen tubing.
• the centrifuge bag is removably fixed within the centrifuge rotor by inserting the raised column through the bag center core and securing with the cover. During the rotation of the centrifuge, components of the whole blood will assume a radial, horizontal position within the centrifuge bag based upon a density of such components, and thus the fluid medium components will be separated from other components having different densities.
• the present invention provides for the specific removal of the desired fraction within one or more of the regions from the centrifuge bag through the outlet tube during continued rotation ofthe centrifuge, thereby allowing for on-line removal ofthe desired fraction. Additional aliquots may be added to the centrifuge bag via the inlet tube simultaneously or after the desired component has been harvested.
• the centrifuge bag is a flexible, transparent, generally flat doughnut-shaped bag.
• the centrifuge bag is a rigid, transparent container having an interior chamber for receiving and holding the fluid medium during centrifugation, the interior chamber having a generally off-centered figure eight cross-sectional configuration.
• Another aspect of the present invention comprises a disposable centrifuge bag having an inlet tube and an outlet tube, wherein the outlet tube is fluidly connected with a bent fitting.
• centrifuge rotor for holding a centrifuge bag, the rotor comprising a base and a cover, the base further having a first grooved, raised center column and the cover having a second grooved, raised center column.
• the centrifuge bag is a flexible, doughnut-shaped bag comprising inlet and outlet tubes in fluid communication with the environment inside and outside the centrifuge bag, wherein the tubes are seated in the base and cover column grooves to hold the centrifuge bag in a fixed position relative to the base and cover, such that the bag does not spin independently ofthe base and cover but rather spins concurrently and at the same rate of rotation as the base and cover.
• Another aspect ofthe present invention comprises a centrifuge rotor for holding a centrifuge bag, the rotor comprising a base and a cover for securing a centrifuge bag therebetween, the centrifuge cover further comprising one or more concentric indicator circles that are spaced from the center ofthe cover or the base to aid the operator in visualizing the distal ends of these tubes.
• centrifuge rotor comprising an interior chamber having a complex configuration, wherein the chamber holds a flexible, doughnut-shaped centrifuge bag for retaining the fluid medium during centrifugation.
• the centrifuge rotor is defined by a base having a lower chamber, and a cover having an upper chamber. When the cover is superimposed on the base, the upper and lower chambers define the annular interior chamber ofthe rotor.
• the interior rotor chamber has a generally off-centered figure eight-shaped cross-sectional configuration specifically designed to maximize the collection of the desired component (e.g., platelet rich plasma) by centrifugation of a fluid medium (e.g., anticoagulated whole blood).
• the centrifuge bag is formed from a substantially flexible material, such that the profile of the centrifuge bag during centrifugation is thus determined at least in part by the volume of the fluid medium contained therein. When the centrifuge bag is filled to maximum capacity, it assumes the configuration ofthe interior ofthe rotor chamber.
• Another aspect of this invention comprises a method for on-line harvesting of a predetermined component of a fluid medium.
• One embodiment ofthe present invention utilizes a centrifuge and a disposable centrifuge bag for containing the fluid medium during separation and which is positionable within the centrifuge, the centrifuge bag further comprising at least one inlet tube and at least one outlet tube.
• the centrifuge includes a centrifuge rotor having a base portion, a cover, and an outer rim. The base portion and the cover define the interior of the centrifuge rotor, which is separated into upper and lower chambers.
• the disposable centrifuge bag is positionable horizontally within the lower chamber and may be appropriately secured to the centrifuge base by the cover.
• the centrifuge bag is fluidly connected via a dual lumen tubing to a source (e.g., to a container comprising anticoagulated autologous whole blood) and collection container ( e -g-, for receiving platelet rich plasma or some other component that will then be further processed).
• the dual lumen tubing comprises an inlet lumen fluidly connected to the inlet tube ofthe centrifuge bag and an outlet lumen fluidly connected to the outlet tube of the centrifuge bag.
• the centrifuge bag is substantially annular relative to the rotational axis of the centrifuge.
• the fluid medium may be provided to the centrifuge bag via the inlet lumen of the tubing during rotation ofthe centrifuge.
• the components of the bag assume radial, horizontal positions base based on their densities.
• the desired fraction may be removed from the centrifuge bag via the outlet lumen during continued rotation ofthe centrifuge.
• the position ofthe fraction to be harvested may be shifted into the area of the outlet tube as needed, either by withdrawing components that are positioned near the outer perimeter through the inlet tube, or by adding additional aliquots of the fluid medium to the bag.
• the bag is a flexible, transparent doughnut-shaped bag.
• the bag is a rigid, transparent bag comprising an interior chamber having an off-centered, figure eight cross- sectional configuration.
• a further object ofthe present invention is to provide for a method and device for the production and isolation of thrombin for all medical uses.
• a further object ofthe present invention is to provide an autologous platelet gel for any application.
• Figure 1 is a perspective view illustrating one embodiment of the continuous-flow centrifugal processing system ofthe present invention illustrating a centrifuge and side- mounted motor positioned within a protective housing or enclosure ofthe invention.
• FIG 2 is an exploded side view ofthe centrifuge and the side-mounted motor of the centrifugal processing system of Figure 1 illustrating the individual components ofthe centrifuge.
• Figure 3 is a partial perspective view of the lower case assembly ofthe drive shaft assembly of Figure 2.
• Figure 4 is an exploded side view ofthe lower case assembly of Figure 3.
• Figure 5 is an exploded perspective view ofthe components of the lower case assembly of Figure 3.
• Figure 6 is a top view of the lower bearing assembly which is positioned within the lower case assembly of Figure 3.
• Figure 7 is a perspective view of the lower bearing assembly of Figure 6.
• Figure 8 is an exploded side view ofthe lower bearing assembly of Figures 6 and
• Figure 9 is a perspective view of the receiving tube guide ofthe centrifuge of Figure 2.
• Figure 10 is an exploded, perspective view of a gear of the mid-shaft gear assembly of Figure 2.
• Figure 11 is a perspective view ofthe gear of Figure 10 as it appears assembled.
• Figure 12 is an exploded, perspective view of the top bearing assembly of the centrifuge of Figure 2.
• Figure 13 is a perspective view of the top case shell of the top bearing assembly of Figure 12.
• Figure 14 is a perspective view of the centrifuge ofthe present invention shown in Figure 1, having a quarter section cut away along lines 14-14 of Figure 1.
• Figure 15 is a perspective view of one embodiment of a centrifuge rotor base.
• Figure 16 is a perspective view of one embodiment of a centrifuge rotor cover.
• Figure 17 is a side cross-sectional view of one embodiment of a rotor of this invention taken along view lines 17 of Figure 14 for holding a disposable centrifuge bag, showing a dual lumen tubing connected to the bag.
• Figure 18 is a side cross-sectional view of one embodiment of a rotor of this invention taken along view lines 18 of Figure 1 for holding a disposable centrifuge bag, showing the grooved columns ofthe base and cover.
• Figure 19 is an enlarged perspective view similar to Figure 1 illustrating an alternate embodiment of a centrifuge driven by a side-mounted motor (with only the external drive belt shown).
• Figure 20 is a cutaway side view ofthe centrifuge of Figure 19 illustrating the internal pulley drive system utilized to achieve a desired drive ratio and illustrating the rotor base configured for receiving a centrifuge bag.
• Figure 21 is a cutaway side view similar to Figure 20 with the rotor base removed to better illustrate the top pulley and the location of both idler pulleys relative to the installed internal drive belt.
• Figure 22 is a sectional view ofthe centrifuge of Figure 20 further illustrating the internal pulley drive system an showing the routing of the centrifuge tube (or umbilical cable).
• Figure 23 is a top view of a further alternate centrifuge similar to the centrifuge of Figure 19 but including internal, separate bearing members (illustrated as four cam followers) that allows the inclusion of guide shaft to be cut through portions of the centrifuge for positioning ofthe centrifuge tube (or umbilical cable).
• internal, separate bearing members illustrated as four cam followers
• Figure 24 is a perspective view similar to Figure 19 illustrating the centrifuge embodiment of Figure 23 further illustrating the guide slot and showing that the centrifuge can be driven by an external drive belt.
• Figure 25 is a top view of a flexible, disposable centrifuge bag of this invention.
• Figure 26 is a perspective view of a flexible, disposable centrifuge bag of this invention.
• Figures 27, 28, 29, and 30 are illustrations of bent fittings of this invention having "T” shaped, “curved T” shaped, “L” shaped, and “J” shaped configurations, respectively.
• Figure 31 is an illustration of an inlet and/or outlet tube of this invention.
• Figure 32 is a top view of a disposable centrifuge bag of this invention after the centrifugation of whole blood, showing the separated blood components.
• Figures 33-39 are schematic illustrations of one method of this invention for separating whole blood components using a disposable centrifuge bag of this invention.
• Figure 40 is a top view of an alternate embodiment of a disposable centrifuge bag ofthe present invention having inner and outer chambers.
• Figure 41 is a top view ofthe disposable centrifuge bag shown in Figure 34 illustrating movement ofthe red blood cell layer from the outer perimeter toward the inner perimeter.
• Figure 42 is a bottom view of an alternate embodiment of a disposable centrifuge bag of the present invention having inner and outer chambers in fluid communication with outlet and inlet ports.
• Figure 43 is a side cross-sectional view of a rigid disposable centrifuge bag of this invention.
• Figure 44 is a schematic illustration of separated blood components contained in a centrifuge bag having an elliptical cross-sectional view of the centrifuge bag shown in Figure 43.
• Figure 45 is a side cross-sectional view of a rigid disposable centrifuge bag of this invention.
• Figure 46 is a schematic illustration ofthe surface areas and various dimensions of the figure eight configuration as shown in Figure 45.
• Figure 47 is a schematic illustration of separated blood components contained in a centrifuge bag having a figure eight side cross-sectional configuration.
• Figure 48 is a side cross-sectional view of an alternative embodiment of an assembled centrifuge rotor of this invention comprising the rotor cover of Figure 49 and the rotor base of Figure 50.
• Figure 49 is a side cross-sectional view of an alternative embodiment of a rotor cover of this invention.
• Figure 50 is a side cross-sectional view of an alternative embodiment of a rotor base of this invention.
• Figure 51 is a perspective view of the rotor base of Figure 50.
• Figure 52 is a perspective view of the rotor cover of Figure 49.
• Figure 53 is a block diagram illustrating the components of a centrifugal processing system of the present invention.
• Figure 54 is a graph illustrating the timing and relationship of transmission of control signals and receipt of feedback signals during operation of one embodiment of the automated centrifugal processing system of Figure 53.
• Figure 55 is a side view of an alternative embodiment of the automated centrifugal processing system of Figure 53 showing a centrifuge having a rotor wherein the reservoir extends over the outer diameter ofthe centrifuge portion that facilitates use of an externally-positioned sensor assembly.
• Figure 56 is a side view of a further alternative embodiment ofthe external sensor assembly feature ofthe centrifugal processing system ofthe invention without an extended rotor and illustrating the positioning of a reflector within the centrifuge.
• Figure 57 is a side view of yet another embodiment ofthe external sensor assembly feature ofthe centrifugal processing system ofthe invention illustrating a single radiant energy source and detector device. ⁇ .
• Figure 58 is a block diagram of a an automated centrifugal processing system, similar to the embodiment of Figure 47, including components forming a temperature control system for controlling temperatures of separated and processed products.
• Figure 59 is a perspective view of components of the temperature control system of
• Figure 60 is schematic and sectional view ofthe dispenser of the present invention.
• Figure 61 is a flow diagram representing the method for isolating platelet rich plasma and platelet poor plasma for use in preparing a platelet gel ofthe present invention.
• Figure 62 is a flow diagram representing the final portion ofthe method for preparing a platelet gel of the present invention using platelet rich plasma as a starting material.
• Figure 63 is a flow diagram representing the final portion ofthe method for preparing a platelet gel ofthe present invention using platelet poor plasma as a starting material.
• Figure 64 is a graphic representation of the effect that the serum-to-plasma ratio has on clotting times.
• Figure 65 graphically represents the effect of calcium addition on the clotting times of platelet rich plasma and platelet poor plasma.
• Figure 66 is a graphic representation ofthe relationship between clotting time and actual gel time using blood drawn from a donor.
• Figure 67 is a graphic representation ofthe relationship between clotting time and actual gel time using blood drawn from a donor.
• Figure 68 graphically represents the effect of calcium addition on clotting times and gel times using blood drawn from a donor.
• Figure 69 graphically represents the effect of calcium addition on clotting times and gel times using blood drawn from a donor.
• centrifugal processing system 10 of the present invention is best shown in
• FIG. 1 having a stationary base 12, a centrifuge 20 rotatably mounted to the stationary base 12 for rotation about a predetermined axis A, a rotor 202 for receiving a disposable bag (not shown) designed for continuous-flow.
• the centrifugal processing system 10 includes a protective enclosure 11 comprising the main table plate or stationary base 12, side walls 13, and a removable lid 15 made of clear or opaque plastic or other suitable materials to provide structural support for components of the centrifugal processing system 10, to provide safety by enclosing moving parts, and to provide a portable centrifugal processing system 10.
• the centrifugal processing system 10 further includes a clamp 22 mounted over an opening (not shown) in the lid 15.
• Clamp 22 secures at a point at or proximately to axis A without pinching off the flow of fluid that travels through umbilical cable 228.
• a side mounted motor 24 is provided and connected to the centrifuge 20 by way of a drive belt 26 for rotating the drive shaft assembly 28 (see Figure 2) and the interconnected and driven rotor assembly 200 in the same rotational direction with a speed ratio selected to control binding of umbilical cable 228 during operation of the system, such as a speed ratio of 2:1 (i.e., the rotor assembly 200 rotates twice for each rotation ofthe drive shaft assembly 28).
• the present invention is further directed toward a dispensing device 902, best shown in Figure 60 for the withdrawal and manipulation of specific blood components for various therapeutic regimens, such as but not limited to the production of platelet rich plasma, platelet poor plasma, and white blood cells which may be used for the production of autologous thrombin and autologous platelet gels.
• the continuous-flow centrifugal processing system 10 comprises a centrifuge 20 to which a rotor 202 is removably or non-removably attached.
• centrifuge 20 and its self-contained mid-shaft gear assembly 108 (comprised of gears 110, 110', 131, and 74) is a key component ofthe invention thereby allowing for the compact size of the entire centrifugal processing system 10 and providing for a desired speed ratio between the drive shaft assembly 28 and the rotor assembly 200.
• the centrifuge 20 is assembled, as best seen in Figure 2, by inserting the lower bearing assembly 66 into lower case shell 32 thus resulting in lower case assembly 30. Cable guide 102 and gears 110 and 110' are then positioned within lower case assembly 30, as will be discussed in more detail below, so that gears 110 and 110' are moveably of engaged with lower bearing assembly 66. Upper bearing assembly 130 is then inserted within top case shell 126 thus resulting in bearing assembly 124 which is then mated to lower case assembly 30, such that gears 110 and 110' are also moveably engaged with upper bearing assembly 130, and held in place by fasteners 29. Lower bearing assembly 66 is journaled to stationary base or main table plate 12 by screws 14, thus allowing centrifuge 20 to rotate along an axis A, pe ⁇ endicular to main table plate 12 (as shown in
• the lower case assembly 30 is preferably, but not necessarily, machined or molded from a metal material and includes a lower case shell 32, timing belt ring 46, timing belt flange 50, and bearing 62 (e.g., ball bearings and the like).
• Lower case shell 32 includes an elongated main body 40 with a smaller diameter neck portion 36 extending from one end ofthe main body 40 for receiving timing belt ring 46 and timing belt flange 50.
• the larger diameter main body 40 terminates into the neck portion 36 thereby forming an external shoulder 38 having a bearing surface 42 for timing belt ring 46.
• Timing belt ring 46 and timing belt flange 50 have inner diameters that are slightly larger than the outer diameter of neck portion 36 allowing both to fit over neck portion 36.
• Shoulder 38 further contains at least one and preferably four internally thread holes 44 that align with hole guides 48 and 52 in timing belt ring 46 and timing belt flange 50, respectively (shown in Figure 5). Consequently, when assembled, screws 54 are received by hole guides 52 and 48 and are threaded into thread holes 44 thus securing timing belt 46 and timing belt flange 50 onto neck portion 36.
• Lower case shell 32 also has an axial or sleeve bore 56 extending there through, and an internal shoulder 58, the upper surface 60 of which is in approximately the same horizontal plane as external shoulder 38.
• Bearing 62 (shown in Figure 4) is press fit concentrically into sleeve bore 56 so that it sits flush with upper surface 60.
• Internal shoulder 58 also has a lower weight bearing surface 64 which seats on the upper surface
• Lower bearing assembly 66 comprises a lower gear insert 70, ball bearings 84, gear 74 and spring pins 76 and 76'.
• the gear 74 may be of any suitable gear design for transferring an input rotation rate to a mating or contacting gear, such as the gears 110, 110' ofthe mid-shaft gear assembly 108, with a size and tooth number selected to provide a desired gear train or speed ratio when combined with contacting gears.
• the gear 74 may be configured as a straight or spiral bevel gear, a helical gear, a worm gear, a hypoid gear, and the like out of any suitable material.
• the gear 74 is a spiral gear to provide a smooth tooth action at the operational speeds ofthe centrifugal processing system 10.
• the upper surface 68 of lower gear insert 70 comprises an axially positioned sleeve 72, which receives and holds gear 74.
• gear 74 is preferably retained within sleeve 72 by the use of at least one and preferably two spring pins 76 and 76' which are positioned within spring pin holes 73 and 73' extending horizontally through lower gear insert 70 into sleeve 72.
• gear 74 having spring pin receptacles 77 and 77' when gear 74 having spring pin receptacles 77 and 77' is inserted into sleeve 72 the spring pins 76 and 76' enter the corresponding receptacles 77 and 77' thus holding the gear 74 in place.
• gear 74 may be held in sleeve 72 by a number of other methods, such as, but not limited to being press fit or frictionally fit, or alternatively gear 74 and lower gear insert 70 may be molded from a unitary body.
• the base 78 of lower gear insert 70 has a slightly larger diameter than upper body 80 of lower gear insert 70 as a result of a slight flare. This slight flare produces shoulder 82 upon which ball bearing 84 is seated.
• a second retaining ring 87 (shown in Figure 2) is also inserted into the annular space produced by the difference between the outer diameter of the lower bearing assembly 66 and the inner diameter of sleeve bore 56 below ball bearing 84, thereby securing lower gear insert 70 within lower case shell 32. Consequently, ball bearings 62 and 84 are secured by retaining rings 86 and 87, respectively, resulting in lower case shell 32 being journaled for rotation about lower bearing assembly 66 but fixed against longitudinal and transverse movement thereon. Therefore, when assembled lower bearing assembly 66 is mounted to stationary base 12, by securing screws 14 into threaded holes 79 located in the base 78. Lower case shell 32 is thus able to freely rotate about stationary lower bearing assembly 66 when the drive belt 26 is engaged.
• tube guide 102 extending from the opposite end of neck portion 36 on lower case shell 32 are a number of protrusions or fingers 88, 90, 92, and 94. Positioned between protrusions 88 and 90, and between protrusions 92 and 94 are recessed slots 96 and 98, respectively, for receiving tube guide 102 ( Figure 9).
• tube guide 102 guides umbilical cable 228 connected to centrifuge bag 226 through the mid-shaft gear assembly 108 and out of the centrifuge 20.
• gears 110 and 110' are preferably configured to provide mating contact with the gear 74 and to produce a desired, overall gear train ratio within the centrifuge 20.
• the gears 110 and 110' are preferably selected to have a similar configuration (e.g., size, tooth number, and the like) as the gear 74, such as a spiral gear design.
• mid-shaft gear assembly 108 comprises a pair of gears 110 and 110'engaged with gears 74 and 131. While the construction of gears and gear combinations is well known to one skilled in the mechanical arts, a brief description is disclosed briefly herein.
• Figure 10 illustrates an exploded view depicting the assembly of gear 110
• Figure 11 is a perspective view of the gear 110 of Figure 10 as it appears assembled.
• gear 110' is constructed in the same manner. Gear 111 is locked onto mid-gear shaft 112 using key stock 114 and external retaining ring 116. Ball bearing 118 is then attached to mid gear shaft 112 using a flat washer 120 and cap screw 122. Recessed slots 104 and 106 of lower case shell 32 then receive ball bearing 118 and 118' (not shown). In an alternate embodiment ball bearing 118 can be replaced by bushings (not shown).
• gears 110 and 110' make contact with the lower gear 74 (see Figures 2 and 14) to provide contact surfaces for transferring a force from the stationary gear 74 to the gears 110 and 110' to cause the gears 110 and 110' to rotate at a predetermined rate that creates a desired output rotation rate for the driven rotor assembly 200.
• the rotor assembly 200 is driven by the drive shaft assembly 28 which is rotated by the drive motor 24 at an input rotation rate or speed, and in a preferred embodiment, the drive shaft assembly 28 through the use of the gears 110 and 110' is configured to rotate the rotor assembly 200 at an output rotation rate that is twice the input rotation rate (i.e., the ratio ofthe output rotation rate to the input rotation rate is 2: 1).
• This ratio is achieved in the illustrated embodiment by locking the gears 110 and 110' located within the drive shaft assembly 28 to rotate about the centrifuge center axis, A, with the lower case shell 32 which is rotated by the drive motor 24.
• the gears 110 and 110' also contact the stationary gear 74 which forces the gears 110, 110' to rotate about their rotation axes which are traverse to the centrifuge center axis, A, and as illustrated, the rotation axes of the gears 110, 110' coincide.
• the gears 110, 1 10' are able to provide the desired input to output rotation rate of 2:1 to the rotor assembly 200.
• Top bearing assembly 124 (as shown in Figure 12) comprises top case shell 126, ball bearing 128, and an upper bearing 130.
• Top case shell 126 as best seen in Figures 12 and 13, comprises an upper surface 132, a lower lip 134 and a central or axial bore 136 there through.
• Upper surface 132 slightly overhangs axial bore 136 resulting in a shoulder 138 having a lower surface 140 (shown in Figure 13).
• Lower lip 134 is a reverse image of upper lip 100 on lower case shell 32 ( shown in Figure 5).
• Upper bearing assembly 130 ( Figure 12) comprises an upper surface 133 and a lower surface 135 wherein the upper surface 133 has a means for receiving a rotor 202. On the lower surface 135 a concentrically positioned column 137 protrudes radially outward pe ⁇ endicular to lower surface 135. Upper bearing assembly 130 further comprises an axially positioned bore 139 that traverses column 137 and upper surface 133 and receives upper gear insert 131. Upper gear insert 131 also contains an axial bore 142 and thus when positioned concentrically within column 137 axial bores 139 and 142 allow for umbilical cable 228 to travel through upper bearing assembly 130 of top case shell 126 down to cable guide 102 (shown in Figure 14).
• upper gear insert 131 may be any suitable gear design for receiving an input rotation rate from a mating or contacting gear, such as the gears 110, 110' ofthe mid-shaft gear assembly 108, with a size and tooth number selected to provide a desired gear train or speed ratio when combined with contacting gears.
• gear insert 131 may be configured as a straight or spiral bevel gear, a helical gear, a worm gear, a hypoid gear, and the like.
• gear 131 is a spiral gear to provide a smooth tooth action at the operational speeds ofthe centrifugal processing system 10.
• Gear insert 131 is preferably retained within column 137 by use of at least one and preferably two spring pins (not shown); however, other assembly techniques may be used to position and retain the gear insert 131 within the column 137 and such techniques are considered within the breadth of this disclosure.
• gear insert 131 may be held in column 137 by a number of other methods, such as, but not limited to being press fit or frictionally fit or alternatively gear insert 131 and the upper bearing assembly may be molded from a unitary body.
• Upper bearing assembly 130 is then inserted into axial bore 136 of top case shell 126 so that the lower surface 135 sits flush with upper surface 132 of top case shell 126.
• Ball bearing 128 is then inserted into the annular space created between the outer diameter of column 137 and the inner side wall 141 of top case shell 126 thereby securing upper bearing assembly 130 into place.
• lower lip 134 is contoured to mate with protrusions 88, 90, 92 and 94 extending from lower case shell 32.
• the outer diameter of lower lip 134 matches the outer diameter ofthe upper end of main body 40 of lower case shell 32 and recesses 144 and 148 receive and retain protrusions 88 and 92 respectively, while recesses 146 and 150 receive and retain protrusions 94 and 88, respectively.
• Holes are placed through each recess and each protrusion so that when assembled, fasteners 152 (shown in Figure 12) can be inserted through the holes thereby fastening the top bearing assembly 124 to the lower case assembly 30.
• fasteners 152 shown in Figure 12
• recessed slots 104' and 106' are recessed slots 104' and 106', respectively, for receiving gears 110 and 110' of mid-shaft gear assembly 108 ( Figure 2 and 14).
• the gears 110 and 110' are preferably configured to provide mating contact with the gear insert 131 and to produce a desired, overall gear train ratio within the centrifuge 20.
• the gears 110 and 110' are preferably selected to have a similar configuration (e.g., size, tooth number, and the like) as the gear 131, such as a spiral gear design.
• recessed slots 96' and 98' exist between recesses 144 and 150 and between recesses 146 and 148, respectively.
• the centrifugal processing system 10 provides a compact, portable device useful for separating blood and other fluids in an effective manner without binding or kinking fluid feed lines, cables, and the like entering and exiting the centrifuge 20.
• the compactness ofthe centrifugal processing system 10 is furthered by the use ofthe entirely contained and interior gear train described above that comprises, at least in part, gear 74, gears 110 and 110', and gear insert 131 ofthe upper bearing 130.
• the gear insert 131 ofthe upper bearing 130 is preferably selected to provide a contact surface(s) with the gears 110 and 110' that transfers the rotation rate ofthe gears 110 and 110' and consequently from gear 74 and to the gear insert 131 ofthe upper bearing 130.
• the gear insert 131 ofthe upper bearing 130 is a spiral gear rigidly mounted within the upper bearing 130 to rotate the rotor assembly 200 and having a design similar to that ofthe spiral gear 74, i.e., same or similar face advance, circular pitch, spiral angle, and the like.
• the gear 74 remains stationary as the lower case shell 32 is rotated about the centrifuge axis, A, at an input rotation rate, such as a rotation rate chosen from the range of 0 ⁇ m to 5000 ⁇ m.
• the gears 110, 110' are rotated both about the centrifuge axis, A, with the shell 32 and by contact with the stationary gear 74.
• the spiral gears 110, 110' contact the gear insert 131 ofthe upper bearing 130 causing the gear insert 131 and connected upper bearing 130 to rotate at an output rotation rate that differs, i.e., is higher, than the input rotation rate.
• gear ratios or train ratios i.e., input rotation rate/output rotation rate
• one embodiment ofthe invention provides for a gear train ratio of 1 :2, where the combination and configuration ofthe gear 74, gears 110, 110', and gear 131 ofthe upper bearing 130 are selected to achieve this gear train ratio.
• the rotation ofthe gears 110, 110' positively affects the achieved gear train ratio to allow, in one embodiment, the use of four similarly designed gears which lowers manufacturing costs while achieving the increase from input to output rotation speeds.
• numerous combinations of gears in differing number, size, and configuration that provides this ratio (or other selected ratios) may be utilized to practice the invention and such combinations are considered part of this disclosure.
• gears 110' are shown in the mid-shaft gear assembly 108 to distribute transmission forces and provide balance within the operating centrifuge, more (or less) gears may be used to transmit the rotation of gear 74 to the gear ofthe upper bearing 130. Also, just as the number, size, and configuration ofthe internal gears may be varied from the exemplary illustration of Figures 1-14, the material used to fabricate the gear 74, the gears 110, 1 10', and the gear insert 131 may be any suitable gear material known in the art.
• the drive motor 24 is mounted on the stationary base 12 of the enclosure 11 adjacent the centrifuge 20.
• the drive motor 24 may be selected from a number of motors, such as a standard electric motor, useful for developing a desired rotation rate in the centrifuge 20 ofthe centrifugal processing system 10.
• the drive motor 24 may be manually operated or, as in a preferred embodiment, a motor controller may be provided that can be automatically operated by a controller of the centrifugal processing system 10 to govern operation of the drive motor 24 (as will be discussed in detail with reference to the automated embodiment ofthe invention).
• a drive belt 26 may be used to rotate the drive shaft assembly 28 (and, therefore, the rotor assembly 200).
• the drive belt 26 preferably has internal teeth (although teeth are not required to utilize a drive belt) selected to mate with the external teeth of the timing belt ring 46 of the lower case assembly 30 portion ofthe drive shaft assembly 28.
• the invention is not limited to the use of a drive belt 26, which may be replaced with a drive chain, an external gear driven by the motor 24, and any other suitable drive mechanisms.
• the drive motor 24 rotates the drive shaft assembly 28 at nearly the same rotation rate (i.e., the input rotation rate).
• a single speed drive motor 24 may be utilized or in some embodiments, a multi and/or variable speed motor 24 may be provided to provide a range of input rotation rates that may be selected by the operator or by a controller to obtain a desired output rotation rate (i.e., a rotation rate for the rotor assembly 200 and included centrifuge bag 226.
• the present invention generally includes an apparatus and methods for the separation of a predetermined fraction(s) from a fluid medium utilizing the principles of centrifugation.
• a predetermined fraction(s) e.g., platelet rich plasma or platelet poor plasma
• the platelet rich plasma may be used, for example, in the preparation of platelet concentrate or gel, and more particularly may be used to prepare autologous platelet gel during surgery using blood drawn from the patient before or during surgery.
• a fluid collection device also referred to as a bowl or rotor 202 is attached to the upper surface 133 ofthe upper bearing assembly 130 as shown in Figures 1 and 2.
• Rotor 202 is preferably mounted permanently to upper bearing assembly 130, however, rotor 202 may also be capable of being removed.
• Rotor 202 comprises a rotor base 204 (shown in Figure 15) having a lower annular groove 212, and a rotor cover 206 having an upper annular groove 214.
• the annular interior chamber 216 of rotor 202 is defined by upper and lower annular grooves 212 and 214.
• the lower annular 212 receives a centrifuge bag 226 for containing the fluid medium to be centrifuged.
• Centrifuge bag 226 is connected to supply and receiving containers 398, 400, respectively, via umbilical cable 228 which is preferably, but not limited to a dual lumen.
• umbilical cable 228 which is preferably, but not limited to a dual lumen.
• umbilical cable 228 may comprise multiple lumens.
• Umbilical cable 228 according to the preferred embodiment comprises inlet lumen 230 and outlet lumen 232 such that a fluid medium may be provided to and removed from the centrifuge bag 226 during rotation of the centrifuge rotor 202.
• centrifuge rotor 202 is more particularly illustrated in Figures 15, 16, 17 and 18.
• Figure 15 is a perspective view of rotor base 204
• Figure 16 is a perspective view of rotor cover 206.
• Figure 17 is a cross-sectional side view of rotor 202 taken along view lines 17 in Figure 1
• Figure 18 is a cross-sectional side view of rotor 202 taken along view lines 18 in Figure 1.
• rotor base 204 comprises raised annular rim 208 and raised column 218 that is axially disposed in base 204.
• Raised column 218 further has a groove 222 extending across the diameter of column 218.
• Annular groove 212 is defined by raised annular rim 208 and raised column
• Rotor cover 206 shown in Figure 16 comprises raised annular rim 210 and raised column 220 which is axially disposed in rotor cover 206.
• Raised column 220 further has a groove 224 extending across the diameter of column 220.
• Annular groove 214 is defined by rim 210 and column 220.
• the height of rim 210 is equal to the height of column 220.
• a flexible centrifuge bag such as a doughnut-shaped centrifuge bag 226 ( Figure 19 and 20) having a center core 242 is placed in rotor base 204 such that center column 218 extends through the core 242 of centrifuge bag 226 and the centrifuge bag 226 lies in annular groove 212.
• Rotor cover 206 is superimposed on rotor base 204 such that grooves 222 and 224 are aligned, as illustrated in Figures 17 and 18.
• rims 208 and 210 are in complete contact with each other such that annular groove 212 and annular groove 214 define rotor interior chamber 216.
• columns 218 and 220 are in complete contact with each other.
• the inner perimeter 240 of centrifuge bag 226 is secured between columns 218 and 220 such that columns 218 and 220 do not completely physically contact each other.
• centrifuge 640 utilizes a uniquely arranged internal pulley system to obtain a desired input to output drive ratio (such as 2:1, as discussed above) rather than an internal gear assembly.
• the centrifuge 640 utilizes the side-mounted motor 24 (shown in Figure 1) through drive belt 26 to obtain the desired rotation rate at the rotor portion ofthe centrifuge.
• the centrifuge 640 includes a rotor base 644 (or top plate) with a recessed surface 648 for receiving and supporting a centrifuge bag during the operation ofthe centrifuge 640.
• the rotor base 644 is rigidly mounted with fasteners (e.g., pins, screws, and the like) to a separately ratable portion (i.e., a top pulley 698 discussed with reference to Figures 20 and 21) of a lower case shell 660.
• a cable port 656 is provided centrally in the rotor base 644 to provide a path for a centrifuge tube or umbilical cable that is to be fluidically connected to a centrifuge bag positioned on the recessed surface 648 ofthe rotor base 644.
• the lower case shell 660 includes a side cable port 662 for the umbilical cable to enter the centrifuge 640, which, significantly, the side cable port 662 is located between idler pulleys 666, 668 to provide a spacing between any inserted tube or cable and the moving drive components of the centrifuge 640.
• Idler shaft or pins 664 are mounted and supported within the lower case shell 660 to allow the pins 664 to physically support the pulleys 666, 668.
• the idler pulleys 666, 668 are mounted on the pins 664 by bearings to freely rotate about the central axis ofthe pins 664 during operation of the centrifuge 640.
• the idler pulleys 666, 668 are included to facilitate translation ofthe drive or motive force provided or imparted by the drive belt 26 to the lower case shell 660 to the rotor base 644, as will be discussed in more detail with reference to Figures 20 and 21, and to physically support the internal drive belt 670 within the centrifuge 640.
• the drive belt 26 is driven by the side-mounted motor 24 (shown in Figure 1) and contacts the lower case shell 660 to force the lower case shell 660 to rotate about its central axis.
• the lower case shell 660 is in turn mounted on the base 674 in a manner that allows the lower case shell 660 to freely rotate on the base 674 as the drive belt 26 is driven by the side-mounted motor 26.
• the base 674 is mounted to a stationary base 12 (shown in Figure 1) such that the base 674 is substantially rigid and does not rotate with the lower case shell 660.
• the centrifuge 640 is shown with a cutaway view to more readily facilitate the discussion ofthe use ofthe internal pulley assembly to obtain a desired output to input ratio, such as two to one.
• the base 674 includes vibration isolators 676 fabricated of a vibration absorbing material such as rubber, plastic, and the like through which the base 674 is mounted relatively rigidly to the stationary base
• a drive pulley 680 which is rigidly mounted to the lower case shell 660.
• the lower case shell 660 through drive pulley 680 rotates about its center axis (which corresponds to the center axis of the centrifuge 640).
• This rotation rate of the lower case shell 660 can be thought of as the input rotation rate or speed.
• the lower case shell 660 is mounted on the base to freely rotate about the centrifuge center axis with bearings 690 that mate with the base 674.
• the bearings 690 are held in place between the bottom pulley 692 and the base 674, and the bottom pulley 692 is rigidly attached (with bolts or the like) to the base 674 to remain stationary while the lower case shell 660 rotates.
• the illustrated bearings 690 are two piece bearings which allow the lower case shell 660 to rotate on the base 674.
• An internal drive belt 670 is provided and inserted through the lower case shell 660 to contact the outer surfaces of the bottom pulley 692.
• the belt 670 preferably is installed with an adequate tension to tightly mate with the bottom pulley 692 such that f ⁇ ctional forces cause the belt 670 to rotate around the stationary bottom pulley
• the internal drive belt 670 passes temporarily outside the centrifuge 640 to contact the outer surfaces ofthe idler pulleys 666 and 668. These pulleys 666, 668 do not impart further motion to the belt 670 but rotate freely on pins 664.
• the idler pulleys 666, 668 are included to allow the rotation about the centrifuge center axis by lower case shell 660 to be translated to another pulley (i.e., top pulley 698) that rotates about the same axis.
• the idler pulleys 666, 668 provide non-rigid (or ratable) support that assists in allowing the belt 670 to be twisted without binding and then fed back into an upper portion ofthe lower case assembly 660 (as shown clearly in Figures 20 and 21). As the internal drive belt 670 is fed into the lower case assembly 660, the belt 670 contacts the outer surfaces of a top pulley 698.
• the movement of the internal drive belt 670 causes the top pulley 698 to rotate about the centrifuge center axis.
• the idler pulleys 666 and 668 by the nature of their placement and orientation within the centrifuge 640 relative to the pulleys 692 and 698 cause the rotor base 644 to rotate in the same direction as the lower case shell 660.
• the top pulley 698 rotated about the centrifuge center axis at twice the input rotation rate because it is mounted to the lower case shell 660 via bearings 694 (preferably, a two piece bearing similar to bearings 690 but other bearing configurations can be used) which are mounted to the center shaft 686 ofthe lower case shell 660 to frictionally contact an inner surface ofthe top pulley 698. Since the internal drive belt 670 is rotating about the bottom pulley 692 and the idler pulleys 666, 668 are rotating about the centrifuge central axis by drive belt 26, the top pulley 698 is turned about the centrifuge central axis in the same direction as the lower case shell 660 but at twice the rate.
• bearings 694 preferably, a two piece bearing similar to bearings 690 but other bearing configurations can be used
• the drive force ofthe drive belt 26 and the internal drive belt 670 are combined by the components ofthe centrifuge 640 to create the output rotation rate. While a number of output to input drive ratios may be utilized, as discussed previously, a 2:1 ratio is generally preferable, and the centrifuge 640 is preferably configured such that the second, faster rotation rate ofthe top pulley 698 is substantially twice that ofthe lower case shell 660.
• the use of an internal drive belt 670 in combination with two pulleys rotating about the same axis and the structural support for the pulleys within a rotating housing results in a centrifuge that is very compact and that operates effectively at a 2: 1 drive ratio with relatively low noise levels (which is desirable in many medical settings).
• the 2: 1 drive ratio obtained in the top pulley 698 is in turn passed on to the rotor base 644 by rigidly attaching the rotor base 644 to the top pulley 698 with fasteners 652.
• a centrifuge bag placed on the recessed surface 648 ofthe rotor base 644 is rotated at a rate twice that ofthe umbilical cable 228 that is fed into lower case shell 660, which effectively controls binding as discussed above.
• the bearing 694 (one or more pieces) wrap around the entire center shaft 686 ofthe lower case shell 660.
• the rotor base 644 includes the cable port 656 and the center shaft 686 is configured to be hollow to form a center cable guide. This allows an umbilical cable 228 to be fed basically parallel to the centrifuge center axis to the centrifuge bag (not shown).
• the lower case shell 660 includes the side cable port 662 to provide for initial access to the centrifuge 640 and also includes the side cable guide (or tunnel) 684 to guide the cable 228 through the lower case shell 660 to the hollow portion of the center shaft 686.
• the side port 662 and the side cable guide 684 are positioned substantially centrally between the two idler pulleys 666, 668 to position the cable 228 a distance away from the internal drive belt 670 to minimize potential binding and wear.
• the centrifuge 640 illustrated in Figures 19-22 utilizes two piece bearings for both the bottom and top pulleys 692 and 698, respectively, and to provide a path for the umbilical cable 228 a central "blind" pathway (via side cable guide 684, the hollow center of the center shaft 686, and cable ports 656, 662) was provided in the centrifuge 640. While effective, this "blind" pathway can in practice present binding problems as the relatively stiff cable 228 is fed or pushed through the pathway.
• centrifuge embodiment 700 is provided and illustrated in Figures 23 and 24.
• the upper portions ofthe centrifuge 700 include a guide slot between the idler pulleys 666, 668 that enables an umbilical cable 228 to be fed into the centrifuge 700 from the top with the no components to block the view ofthe operator inserting the cable
• the contiguous upper bearing 694 in the centrifuge 640 are replaced with bearing members that have at least one gap or separation that is at least slightly larger than the outer diameter of the cable 228.
• a number of bearing members may be utilized to provide this cable entry gap and are included in the breadth of this disclosure.
• the centrifuge 700 includes a rotor base 702 that is rigidly fastened with fasteners 704 to the top pulley 698 (not shown) to rotate with this pulley at the output rate (e.g., twice the input rate) and to receive and support a centrifuge bag on recessed surface 716.
• the rotor base 702 further includes the cable port 718 which is useful for aligning the center of the bag and cable 228 with the center of the centrifuge
• the rotor base 702 further includes a cable guide slot 712 which as illustrated is a groove or opening in the rotor base 702 that allows the cable 228 to be inserted downward through the centrifuge 700 toward the side cable guide 724 ofthe lower case shell 720.
• the lower case shell 720 also includes a cable guide slot 722 cut through to the top ofthe side cable guide 724.
• the guide slots 712 and 724 are both located in a portion of the centrifuge 700 that is between the idler pulleys 666, 668 to position an inserted cable 228 from contacting and binding with the internal drive belt 670, which basically wraps around 180 degrees ofthe top pulley or lower case shell 720.
• the bearing members 706 are spaced apart and preferably, at least one of these spaces or gaps is large enough to pass through the cable 228 to the center shaft ofthe lower case shell 720.
• four cam followers are utilized for the bearing members 706, although a different number may be employed.
• the cam followers 706 are connected to the top pulley to enable the top pulley to rotate and are connected, also, to the center shaft ofthe lower case shell 720 to rotate with the lower case shell 720.
• the cam followers 706 ride in a bearing groove 710 cut in the lower case shell 720.
• the cable guide slots 712 and 722 are positioned between the two cam followers 706 adjacent the idler pulleys 666, 668, and preferably the guide slots 712, 722 are positioned substantially centrally between the pulleys 666, 668.
• the guide slots 712, 722 are positioned between these cam followers 706 to position the cable 228 on the opposite side ofthe centrifuge 700 as the contact surfaces between the internal drive belt 670 and the top pulley 698 (shown in Figure 20- 22).
• the bag is an integral two stage self balancing disposable design.
• the disposable centrifuge bag 226 has a substantially flat, toroidal- or doughnut-shaped configuration having outer and inner perimeters 238 and 240, respectively, and comprises radially extending upper and lower sheets 234, 236 formed from a substantially flexible material.
• the upper and lower sheets 234, 236 are superimposed and completely sealed together at outer perimeter 238 by a heat weld, rf
• centrifuge bag 226 further comprises an inlet tube 248 sandwiched between upper and lower sheets 234, 236 and extending from the center of core 242 defined by inner perimeter 240 to the outer perimeter 238 and an outlet tube 250 sandwiched between upper and lower sheets 234, 236 and extending from the center of the core 242 to the outer perimeter 238.
• inlet and outlet tubes 248, 250 are thereby sealed therebetween.
• Inlet and outlet tubes 248, 250 are each in fluid communication with the interior of centrifuge bag 226 and the environment outside centrifuge bag 226. The length of outlet tube 250 is shorter than the length of inlet tube 248.
• outlet tube 250 is a straight tube as shown in Figure 31.
• outlet tube 250 includes a bent fitting 252 fluidly connected to the distal end of outlet tube 250 ( Figures 25 and 26).
• the bent fitting 252 may be of any number of configurations, although preferably bent fitting 252 is shaped in the form of a "T", “curved T”, a “J”, or an "L”, as illustrated in Figures 27, 28, 29 and 30, respectively.
• outlet tube 250 and bent fitting 252 may be one contiguous molded unit rather than two connected pieces.
• bent fitting 252 is in the shape of a "T” or a "curved T” as illustrated in Figures 27 and 28, respectively.
• the "T” or "curved T” design of bent fitting 252 ensures that the desired blood component (fraction) will be removed from the sides ofthe bent fitting 252, rather than from a fraction located above or below the bent fitting, as discussed below in detail.
• inlet and outlet tubes 248, 250 are seated in groove 222.
• inlet and outlet tubes 248, 250 are also seated in groove 224.
• Seating inlet and outlet tubes 248, 250 in grooves 222, 224 ensures that centrifuge rotor 202 is held in a fixed position between rotor base 204 and rotor cover 206 such that the centrifuge bag 226 and centrifuge rotor 202 rotate together. That is, the fixed position of centrifuge bag 226 ensures that centrifuge bag 226 will not rotate independently of centrifuge bag 226 during centrifugation.
• Inlet and outlet tubes 248, 250 are fluidly connected at their proximal ends to umbilical cable 228, which in this particular embodiment is a dual lumen tubing connecting centrifuge bag 226 to source and receiving containers 398, 400, respectively, for the introduction and removal of components from the centrifuge bag 226 during centrifugation (see Figure 17).
• Dual lumen tubing 228 comprises inlet lumen 230, which connects inlet tube 248 of centrifuge bag 226 with source container 398, and outlet lumen 232, which connects outlet tube 250 centrifuge bag 226 with receiving container 400.
• the inlet and outlet tubes 248, 250 are adapted at their proximal ends for inserting into the inlet and outlet lumens 230 and 232, respectively.
• connecting means 254 are inserted into the proximal ends of inlet and outlet tubes 248, 250 for connecting the tubes to the inlet and outlet lumens 230, 232 as illustrated in Figure
• one end of umbilical cable 228 must be secured to rotor assembly 200 to prevent itself from becoming twisted during rotation of rotor assembly 200 by the coaxial half-speed rotation of drive shaft assembly 28, which imparts a like rotation with respect to the rotor 202 axis and consequently to the umbilical cable 228 that is directed through cable guide 102. That is, if rotor assembly 200 is considered as having completed a first rotation of 360° and drive shaft assembly 28 as having completed a 180° half- rotation in the same direction, the umbilical cable 228 will be subjected to a 180° twist in one direction about its axis.
• umbilical cable 228 is subjected to a continuous flexture or bending during operation of the centrifugal processing system 10 of the present invention but is never completely rotated or twisted about its own axis.
• FIG. 35 An alternative embodiment of a disposable centrifuge bag of this invention, shown in Figures 35 comprises two or more inlet tubes and/or two or more outlet tubes, wherein the tubes are fluidly connected to a multiple lumen tubing.
• the disposable centrifuge bag 226 is formed from a transparent, substantially flexible material, including but not limited to, polyvinyl chloride, polyethylene, polyurethane, ethylene vinyl acetate and combinations ofthe above or other flexible materials. Based upon the flexibility ofthe centrifuge bag 226, the profile ofthe flexible centrifuge bag 226, shown in Figures 25 and 26, is determined at least in part by the amount of fluid contained therein. The profile of centrifuge bag 226 is further defined by the interior configuration ofthe centrifuge rotor, as discussed below in detail. The ability to manipulate the profile of centrifuge bag 226 based on the interior configuration ofthe centrifuge rotor is utilized at least in part to maximize the volume of fluid medium that can be contained in centrifuge bag 226 during centrifugation, as will be discussed below.
• the fluid or medium to be centrifuged may be contained within source container 300.
• the fluid i.e., whole blood
• the fluid may be withdrawn from the patient during or prior to surgery into source container 398 containing an anticoagulant.
• the anticoagulated whole blood is introduced to centrifuge bag 226 through inlet tube 248 via inlet lumen 230 after the centrifuge bag 226 has been positioned in the centrifuge rotor 202 and rotation thereof is initiated.
• centrifuge bag 226 As discussed above, securing centrifuge bag 226 in centrifuge base 204 in grooves 222, 224 holds the centrifuge bag 226 in a fixed position therebetween, such that the centrifuge bag 226 cannot move independently of the centrifuge rotor 202, and therefore the centrifuge bag 226 and rotor assembly 200 rotate concurrently at the same rate of rotation. Rotation of the centrifuge rotor 202 directs the heavier density constituents ofthe anticoagulated whole blood within the centrifuge bag
• centrifuge cover 206 may further contain concentric index lines to assist the operator in viewing the positions of outlet tube 250 to the RBC plasma interface.
• the location ofthe WBC and platelets can be varied with respect to the red blood cell's and plasma interface. For example, if the ⁇ m is held low (approximately 1 ,000 - 1 JOO, preferably 1 ,500) the plasma and platelets will separate from the RBC layer, as the ⁇ tn's are increased (1,400 - 1J00) the platelets will separate out of the plasma and reside at the plasma to RBC interface in greater concentrations. With increased speeds WBC reside deeper into the RBC pack.
• bent fittings 252 in one embodiment a bent fitting is fluidly connected to the distal end of outlet tube 250. While bent fitting 252 is shown in Figure 32 as having a "T" shape ( Figure 27), this is for illustrative pu ⁇ oses only. Thus, it will be appreciated that bent fitting 252 as shown in Figure 32 could have a number of other configurations, such as those shown in Figures 25-31.
• the design of bent fitting 252 ensures that the desired component is withdrawn (e.g., the platelet rich plasma fraction 260) with less risk of contamination from withdrawing a portion of the adjacent fraction 258. Thus, in one embodiment, the desired fraction is withdrawn when its position overlaps with the position of bent fitting 252.
• the inlet tube 248 may be first used to draw off the red blood cell fraction 256, and when it is desirable to remove the predetermined fraction from the centrifuge bag 226, the predetermined fraction is drawn through bent fitting 252 and outlet tube 250 and directed to receiving container 400 via outlet lumen 232.
• substantially annular regions having constituents of a particular density or range of densities begin to form.
• the separation of whole blood will be discussed, and as shown in Figure 32 four regions are represented, each of which contains a particular type of constituent of a given density or range of densities.
• there may be a given distribution of densities across each of the regions such that the regions may not be sha ⁇ ly defined. Consequently, in practice the regions may be wider (e.g., a larger radial extent) and encompass a range of densities of constituents.
• the first region 256 is the outermost ofthe four regions and contains red blood cells.
• the second region 258 contains white blood cells, which have a lower density than that ofthe red blood cells.
• the third region 260 contains the platelet rich plasma fraction, and the innermost region 262 contains the least dense platelet poor plasma fraction. In one embodiment, it may be desired to harvest the platelet rich plasma fraction in region 260.
• vacuum or suction is provided via outlet lumen 232 to the centrifuge bag 226 to remove a desired portion of region 260. A portion ofthe fraction 260 that is in the area ofthe bent fitting 252 is drawn through bent fitting 252 and into an appropriate one of the collection containers 400 ( Figure 17).
• Figures 33-39 illustrate one method of this invention for the separation of whole blood components, which is a dynamic process.
• Figure 33 shows one portion of the centrifuge bag 226, illustrating the separation of the whole blood components after infusion of an aliquot of whole blood into centrifuge bag 226 and centrifugation for approximately 60 seconds to 10 minutes at a rate of rotation between 0 and 5,000 rpms. It will be understood by those of skill in the art that faster speeds of rotation will separate the blood in a shorter prior of time.
• Figure 33 shows the four separated whole blood fractions, with the denser fractions closer to outer perimeter 238, and the less dense fractions closer to inner perimeter 240.
• hematocrits i.e., the volume of blood, expressed as a percentage, that consists of red blood cells
• centrifuge bag 226 the volume of blood, expressed as a percentage, that consists of red blood cells
• centrifugation of an initial infusion of an aliquot of anticoagulated whole blood will give the profile shown in Figure 33.
• centrifuge bag 226 has a design as shown in Figure 32 wherein bent fitting 252 positioned approximately in the area where a platelet rich plasma fraction 260 is typically found after centrifugation of an aliquot of whole blood.
• concentric indicator lines 205, 207, and 209 (not shown) in the upper surface of rotor cover 206, wherein the concentric lines 205, 207 and 209 correspond approximately with the edges of regions 260, 258, and 256, respectively.
• concentric lines similar to 205, 207 and 209 may be directly imprinted onto the surface of centrifuge bag 226. After centrifugation of an aliquot of blood contained in centrifuge bag 226, a substantial portion ofthe platelet rich plasma fraction 260 is withdrawn from centrifuge bag 226 through bent fitting 252 while centrifuge rotor 202 is still spinning.
• the innermost fraction 262 naturally moves in the direction ofthe outer perimeter 238 due to centrifugal force, as shown in Figure 34.
• the withdrawal of platelet rich plasma fraction 260 is terminated at a point where the platelet poor plasma fraction 262 is close to bent fitting 252 and before any significant portion of platelet poor plasma fraction 262 could be withdrawn through bent fitting 252, as shown in Figure 34. This point can be determined either visually by the operator by volume, or by a sensor, as described below in detail.
• inlet lumen 230 is disconnected from the whole blood source container 398 and connected to a disposal container, after which the remaining fluid in centrifuge bag 226 is evacuated through inlet tube 248 and directed to the disposal container. The inlet lumen is then reconnected to the whole blood source container, and the above-described batch process is repeated as many times as required until the necessary quantity ofthe desired fraction is isolated.
• the above-described process can be performed as a continuous process wherein the step of disconnecting the inlet lumen 230 from the whole blood source 398 can be avoided.
• the continuous process separation of whole blood may be achieve by using a disposable centrifuge bag 226' as illustrated in Figures 39-39 comprising an inlet tube 248 and three outlet tubes 245, 247 and 250, wherein the tubes are connected to an umbilical cable comprising four lumens.
• a disposable centrifuge bag for use in a continuous separation of whole blood comprises inlet tube 248 connected via an inlet lumen to a whole blood source container, a first outlet tube 250 connected to a first outlet lumen that is in turn connected to a platelet rich plasma receiving container, a second outlet tube 245 connected via a second outlet lumen to either a red blood cell receiving container or a waste container and a third outlet tube 247 connected via a third outlet lumen to a platelet poor plasma receiving container.
• Centrifuge bag has the capacity to receive an additional volume (aliquot) of whole blood. Consequently, as shown in Figure 35 infusion of an aliquot of whole blood is reinitiated through first inlet tube 248 with continued centrifugation until the capacity ofthe centrifuge bag 226' is reached. As a result ofthe additional volume of blood, the profile of the blood fractions in centrifuge bag 226' will approximately assume the profile shown in Figure 35. As can be seen in Figure 35, the additional volume of blood results in a shift of the location ofthe blood fractions, such that the platelet rich plasma fraction 260 has shifted back into the area of the bent fitting 252, and the platelet poor plasma fraction 262 has shifted back towards the inner perimeter 240 and away from the vicinity ofthe bent fitting 252. Additional platelet rich plasma 260 can now be removed from centrifuge bag 226' through outlet tube 250 as shown in Figure 35.
• the increased volume of red blood cells now present in centrifuge bag 226' shifts the location of the fractions towards the inner perimeter 240 such that the white blood cell fraction 260 is now in the vicinity ofthe bent fitting 252 as opposed to the desired platelet rich plasma fraction 262.
• centrifuge bag 226' advantageously provides means for shifting the fractions back to the desired locations when the situation shown in Figure 37 arises. That is, second outlet tube 245 serves as an inlet conduit for introduction of whole blood aliquots into centrifuge bag 226', also serves the function of withdrawing fractions that are located close to the outer perimeter 238.
• second outlet tube 245 having its distal end close to outer perimeter 238, can be operated to withdraw a substantial volume of the red blood cell fraction 256, which in turn shifts the location of the remaining fractions 258, 260, 262.
• the withdrawal ofthe red blood cell fraction 256 may be monitored visually by the operator, or by other means such as a sensor.
• the positions ofthe fractions may be shifted by withdrawing the platelet poor plasma fraction 262 through third outlet tube 247, which is connected via a third outlet lumen to a platelet poor plasma receiving container.
• Figure 37 shows that, after withdrawal of a portion ofthe red blood cell fraction 256, the centrifuge bag 226' again has the capacity to receive an additional volume of whole blood for centrifugation. An additional infusion of an aliquot of whole blood through inlet tube 248 into the centrifuge bag 226' of Figure 37 and centrifugation will produce the profile illustrated in Figure 39.
• the above-described steps may be repeated as needed until the desired amount of platelet rich plasma has been harvested. All ofthe above-described steps occur while the centrifuge rotor 202 is spinning.
• the above-described continuous separation method was illustrated in terms of performing the whole blood infusion step and the platelet rich plasma harvesting step sequentially.
• An alternative embodiment involves performing the infusion and harvesting steps substantially simultaneously, that is, the platelet rich plasma fraction is withdrawn at approximately the same time as an additional aliquot of whole blood is being added to the bag.
• This alternate embodiment requires that the centrifuge rotor spin at a rate that results in almost immediate separation ofthe blood components upon infusion of an aliquot of whole blood.
• centrifuge cover 206 may further include one or more concentric indicator circles 205, 207, 209 (shown in Figures 17 and 18) which may be spaced from the center of cover 206 at distances approximately equal to the outer edges of regions 260, 258 256, respectively, to aid the operator in visualizing the positions of these regions with respect to 252.
• Figures 33-39 illustrate one embodiment of how the design of centrifuge bags 226 and 226' permit the general locations ofthe various blood fractions to be shifted to allow for continuous harvesting of a desired blood fraction without the risk of contaminating the harvested blood fraction, and further allow for continual on-line harvesting of a large volume (10 to 5 L's) of blood using a small, portable centrifuge device comprising a 10 cc to 200 cc capacity disposable centrifuge bags 226 and 226'.
• centrifuge bag 226 having inlet tube 248 and outlet tube 250 means that the desired component or fraction will be withdrawn from centrifuge bag
• centrifuge bag 226 offers a significant advantage over conventional centrifuge containers comprising only one tube which serves to both introduce the fluid medium to the container and to withdraw the harvested fraction from the container.
• centrifuge bags 226 and 226' are independent of composition ofthe whole blood to be centrifuged.
• hematocrits i.e., the percent volume of blood occupied by red blood cells
• the profile illustrated in Figure 32 will vary from individual to individual. That is, the width of red blood cell fraction 256 may be wider or narrower, which in turn will result in the platelet rich plasma fraction 260 being positioned further away in either direction from bent fitting 252.
• the design of centrifuge bags 226 and 226' allow the location ofthe desired fraction to be shifted until it is in the region of bent fitting 252.
• centrifuge bag 270 has a substantially flat, toroidal- or doughnut-shaped configuration having outer and inner perimeters 271 and 272, respectively, and comprises radially extending upper and lower sheets 273, 274 formed from a substantially flexible material.
• the upper and lower sheets 273, 274 are superimposed and completely sealed together at outer perimeter 271 by an rf weld, heat weld or other comparable method of adhering two surfaces.
• Inner perimeter 272 defines core 275 of centrifuge bag 270.
• centrifuge bag 270 further comprises inlet tube 276 sandwiched between upper and lower sheets 273, 274 and radially extending from the center of core 275 to the outer perimeter 271 , and outlet tube
• Inlet and outlet tubes 276, 278 are each in fluid communication with the interior of centrifuge bag 270 and the environment outside centrifuge bag 270.
• Inlet tube 276 and outlet tube 278 are fluidly connected to umbilical cable 228 (not shown), which in this particular embodiment is a dual lumen tubing.
• Inlet tube 276 is fluidly connected at its proximal end to umbilical cable 228, preferably by an L-shaped connector (not shown), and outlet tube 278 is fluidly connected at its center to umbilical cable 228 via a T-shaped connector (not shown).
• the disposable centrifuge bag 270 is formed from a transparent, substantially flexible material, including but not limited to, polyvinyl chloride, polyethylene, polyurethane, ethylene vinyl acetate and combinations ofthe above or other flexible materials. Upper and lower sheets 273, 274 of centrifuge bag 270 are further sealed at two portions between the outer perimeter and the inner perimeter.
• centrifuge bag 270 further comprises a first C-shaped seal 282 located between the outer and inner perimeters 271, 272 and having an first concave indentation or well 283 on the concave side of C- shaped seal 282, and a second C-shaped seal 284 located between the outer and inner perimeters 271, 272 and having an second concave indentation or well 285 on the concave side of C-shaped seal 284.
• First and second C-shaped seals 282 and 284 are formed by sealing portions of upper and lower sheets 273, 274 together by methods known in the art for sealing two surfaces, including but not limited to rf or heat welding.
• first and second C-shaped seals 282, 284 have their concave sides facing each other such that the first and second indentations 283, 285 are diametrically opposed to each other. That is, when centrifuge bag 270 is viewed from the top as in Figure 40, first and second C-shaped seals 282, 284 are mirror images of each other.
• First and second C-shaped seals 282, 284 together define an outer chamber 292 between the outer perimeter 271 and first and second C-shaped seals 282, 284, wherein the outer chamber 292 has a toroidal configuration and serves as a first processing compartment.
• First and second C-shaped seals 282, 284 together further define an inner chamber 293 between first and second C- shaped seals 282, 284 and inner perimeter 272, wherein the inner chamber 293 has a toroidal configuration and serves as a second processing compartment.
• the first and second C-shaped seals 282, 284 are positioned such that ends 288 and 290 are directly opposite and spaced apart from each other to define a first channel 286 therebetween, and such that ends 289 and 291 are directly opposite and spaced apart from each other to define a second channel 287 therebetween, wherein the first and second channels 286, 287 are diametrically opposed and provide fluid communication between the first processing compartment 292 and the second processing compartment 293.
• Inlet tube 276 extends through either channel 286 or channel 287, and the first and second distal ends 280, 281 of outlet tube 278 extend into first and second indentations 283, 285, respectively.
• Centrifuge bag 270 is removably secured between rotor base 204 and rotor cover 206 of rotor 202 in a manner as described above so that centrifuge bag 270 is held in a fixed position relative to rotor base 204 and rotor cover 206 during rotation of the centrifuge rotor 202.
• rotor base 204 Figure 15
• rotor cover 206 Figure 16
• an alternate embodiment of rotor base 204 comprises raised column 218 comprising first and second grooves which are pe ⁇ endicular to each other and extend the diameter ofthe raised base column 218, such that when rotor 202 is assembled, inlet tube 276 and outlet tube 278 of centrifuge bag 270 are seated in the first and second grooves, respectively, of raised base column 218.
• an alternate embodiment of cover 206 comprises raised column 220 comprising first and second grooves which are pe ⁇ endicular to each other and extend the diameter ofthe raised cover column 220, such that when rotor 202 is assembled, inlet tube 276 and outlet tube 278 are further seated in the first and second grooves, respectively, of raised cover column 220.
• inlet and outlet tubes 276, 278 are fluidly connected to umbilical cable 228, which in this particular embodiment is a dual lumen tubing connecting centrifuge bag 270 to source and receiving containers 398, 400, respectively, for the introduction ofthe fluid to be centrifuged in bag 270 and for the removal of one or more of the separated components from the centrifuge bag 270 during rotation of the centrifuge 20.
• Dual lumen tubing 228 comprises inlet lumen 230, which connects inlet tube 276 with source container 398, and outlet lumen 232, which connects outlet tube 278 with receiving container 400.
• the fluid or medium to be centrifuged using centrifuge bag 270 may be contained within source container 398.
• the fluid i.e., whole blood
• source container 398 containing an anticoagulant containing an anticoagulant
• the anticoagulated whole blood is introduced to centrifuge bag 270 through inlet tube 276 via inlet lumen 230 after the centrifuge bag 270 has been positioned in the centrifuge rotor 202 and rotation thereof is initiated.
• Centrifuge bag 270 may be used for the separation and isolation of one or more components dissolved or suspended in a variety of fluid media, including, but not limited to, the separation of cellular components from biological fluids.
• centrifuge bag 270 is useful for the concentration and removal of platelets from whole blood.
• the separation of a fluid medium such as whole blood in centrifuge bag 270 may be considered to be a two-stage separation process.
• the first stage ofthe separation of platelets from whole blood involves separation of a platelet suspension from the red blood cells.
• the platelet suspension is typically plasma rich in platelets, and it is commonly referred to as platelet-rich plasma (PRP).
• PRP platelet-rich plasma
• the term "platelet suspension” is not limited to PRP in the technical sense, but is intended to encompass any suspension in which platelets are present in concentrations greater than that in whole blood, and can include suspensions that carry other blood components in addition to platelets.
• the second stage of the separation comprises separating platelets from the platelet suspension to produce a platelet concentrate.
• the term "platelet concentrate” is intended to encompass a volume of platelets that results after a "platelet suspension” undergoes a subsequent separation step that reduces the fluid volume of the platelet suspension.
• the platelet concentrate may be a concentrate that is depleted of white blood cells and red blood cells.
• stage one of a whole blood separation process using centrifuge bag 270 begins with the introduction of an aliquot of whole blood into centrifuge bag 270 via inlet tube 276 during rotation ofthe centrifuge 20.
• the aliquot of whole blood enters outer chamber 292 of centrifuge bag 270, it quickly separates radially under the influence of centrifugal force into various fractions within outer chamber 292 based on the densities ofthe components ofthe whole blood, including an outermost fraction containing the red blood cells which pack along the outer perimeter 271 of centrifuge bag 270, and an inner fraction comprising the platelet suspension.
• the platelet suspension after centrifugation ofthe first aliquot of whole blood is represented in Figure 41 by ring 294.
• red blood cells are near the entrance of channels 286, 287 may be monitored either visually or by a sensor, as described below in detail. At this point the infusion of additional aliquots of whole blood is terminated, and the second stage of the two-stage separation process begins.
• the platelet suspension which was pushed through channels 286, 287 into the second processing compartment 293 flow under the influence of centrifugal force towards positions within the second processing compartment 293 that have the greatest radial distances, that is, towards concave wells 283, 285, where the platelets, being the higher density component ofthe platelet suspension, begin to collect and pack.
• the platelets can then be withdrawn from concave wells 283, 285 through outlet tube 278.
• First and second concave wells 283, 285 act as reservoirs for containing the platelets as they are separated from the platelet suspension in the second stage ofthe separation process.
• inlet lumen 230 is disconnected from the whole blood source container, after which the remaining components in centrifuge bag 270 are evacuated through inlet tube 276 by applying suction to inlet lumen 230 and are directed to a disposable container.
• the inlet lumen 230 is then reconnected to the whole blood source container, and the above-described batch process is repeated as many times as required until the desired quantity of platelets has been harvested.
• Disposable centrifuge bag 320 has a substantially flat, toroidal- or doughnut-shaped configuration having outer and inner perimeters 322 and 324, respectively, and comprises radially extending upper and lower sheets 323, 325 formed from a substantially flexible material.
• the upper and lower sheets 323, 325 are superimposed and completely sealed together at outer perimeter 322 by an rf weld, heat weld or other comparable method of adhering two surfaces.
• Inner perimeter 324 defines core 327 of bag 320.
• Upper and lower sheets 323, 325 of centrifuge bag 320 are further sealed at two portions between the outer perimeter and the inner perimeter.
• centrifuge bag 320 further comprises a first C-shaped seal 326 located between the inner and outer perimeters 322, 324, and a second C-shaped seal 328 located between the inner and outer perimeters 322, 324.
• the first and second C-shaped seals 326, 328 have their concave sides facing each other such that when centrifuge bag 320 is viewed from the top as in Figure 36, first and second C-shaped seals 326 and 328 are mirror images of each other.
• C-shaped seals 326, 328 together define an outer compartment 348 between the outer perimeter 322 and first and second C-shaped seals 326 and 328, wherein the outer compartment 348 has a toroidal configuration.
• First and second C-shaped seals further define an compartment 350 between first and second C-shaped seals 326, 328 and inner perimeter 324, wherein the inner compartment 350 has a doughnut shaped configuration.
• first and second C-shaped seals 326 and 328 are positioned such that ends 330 and 334 of first and second C-shaped seals 326, 328, respectively, are directly opposite and spaced apart from each other, thereby defining first channel 335 therebetween, and such that ends 332 and 336 of first and second seals 326 and 328, respectively, are directly opposite and spaced apart from each other, thereby defining second channel 337 therebetween, wherein the first and second channels 335 and 337 are diametrically opposed.
• First and second channels 335 and 337 provide fluid communication between the outer and inner compartments.
• Centrifuge bag 320 further comprises an inlet port 340, in the lower sheet 325 for introducing fluid into outer compartment 348.
• the inlet port 340 is spaced 90 degrees from channel 335 however it could also be positioned at an angle greater or less than 90 degrees from channel 335.
• Centrifuge bag 320 further comprises first and second outlet ports 344, 346 in lower sheet 325 and positioned within channels 335 and 337 for withdrawing a fluid compartment from centrifuge bag 320.
• centrifuge bag 320 comprises inlet tube 338 secured to the outside surface of upper sheet 323 or lower sheet 325 and radially extending from the center of core 327 towards the outer perimeter 322, wherein inlet tube 338 is fluidly connected at its distal end to inlet port 340.
• Inlet port 340 fluidly connects inlet tube 338 with the outer chamber 348 of centrifuge bag 320.
• Inlet tube 338 is fluidly connected at its proximal end to umbilical cable 228, preferably by an L-shaped connector (not shown).
• centrifuge bag 320 comprises outlet tube 342 secured to the outside surface of upper sheet 323 or lower sheet 325 and extending across the diameter of core 327, wherein one end of outlet tube 342 is fluidly connected to first outlet port 344 and the other end of outlet tube 342 is fluidly connected to second outlet port
• Outlet tube is fluidly connected at its center to umbilical cable 228 via a T-shaped connector (not shown).
• centrifuge bag 320 comprises inlet tube 338 sandwiched between upper and lower sheets 323, 325 and extending radially from the center of core 327 towards outer perimeter 322, wherein inlet tube 328 is fluidly connected at its distal end to inlet port 340, and outlet tube 342 sandwiched between upper and lower sheets 323, 325 and extending across the diameter of core 327, wherein one end of outlet tube 342 is fluidly connected to outlet port 344 and the other end of outlet tube 342 is fluidly connected to outlet port 346.
• inlet and outlet tubes 338, 342 are thereby sealed therebetween.
• Inlet tube 338 and outlet tube 342 are fluidly connected to umbilical cable 228 (not shown), which in this particular embodiment is a dual lumen tubing.
• Centrifuge bag 320 is removably secured between rotor base 204 and rotor cover 206 of rotor 202 in a manner as described above so that centrifuge bag 320 is held in a fixed position relative to rotor base 204 and rotor cover 206 during rotation ofthe centrifuge rotor 202.
• rotor base 204 Figure 15
• rotor cover 206 Figure 16
• Centrifuge bag 370 may be used for the separation and isolation of one or more components dissolved or suspended in a variety of fluid media, including, but not limited to, the separation of cellular components from biological fluids.
• centrifuge bag 370 is useful for the concentration and removal of platelets from whole blood. Therefore, the following description ofthe separation of platelets from whole blood using centrifuge bag 320 is merely for pu ⁇ oses of illustration and is not meant to be limiting of the use of bag 320.
• the separation of a fluid medium such as whole blood in centrifuge bag 320 may be considered to be a one-stage separation process.
• centrifugation of whole blood begins with the introduction of an aliquot of whole blood into centrifuge bag 320 through inlet port 340 via inlet tube 338 during rotation of the centrifuge 20.
• Inlet tube 338 is fluidly connected via inlet lumen 230 of umbilical cable 228 to an anticoagulated whole blood source.
• the aliquot of whole blood enters the outer chamber 348 of centrifuge bag 320, it quickly separates radially within outer chamber 348 into various fractions based on the densities ofthe components of the whole blood, including an outermost fraction containing the red blood cells which pack along the outer perimeter 322 of centrifuge bag 320, and inner fractions containing the platelets and plasma.
• inlet lumen 230 is disconnected from the whole blood source container and connected to a disposal container, after which the remaining components in centrifuge bag 320 are evacuated through inlet port 34- by applying suction to inlet tube 276 and are directed to a disposal container.
• the inlet lumen 230 is then reconnected to the whole blood source container, and the above-described process is repeated as many times as required until the desired quantity of platelets has been harvested.
• a centrifuge container of this invention for the separation of components in a fluid medium (e.g., whole blood), shown in Figures 45-52, is designed to position the desired component (e.g., platelet rich plasma) the platelet rich plasma at a region within the fixed centrifuge container or centrifuge bag so that the desired fraction has a maximum horizontal surface area (i.e., width).
• a centrifuge container 500 shown in Figure 45.
• Figure 45 is a side cross- sectional view of a rigid container 500 comprising a rigid, annular body 510 having an axial core 600 that is closed at the top end 610 and opened at the bottom end 620.
• Rigid container 500 further comprises an interior collection chamber 580 for receiving and holding the fluid medium to be centrifuged and having an outer perimeter 585 and an inner perimeter 590.
• the side, cross-sectional profile of chamber 580 is generally an off- centered "figure eight" or “dumbbell” shape, as shown in Figure 46.
• “figure eight" or “dumbbell” shaped means that the height of section A is approximately equal to the height of section C, and the heights of sections A and C are greater that the height of section B.
• off-center means that the width Wi from the center of section B to outer perimeter 585 is less than the width W 2 from the center of section B to inner perimeter 590 as shown in Figure 45 and 46.
• Rigid container 500 further comprises inlet channel 550 extending radially from core 600 to a point near the outer perimeter 585 and is fluidly connected at its distal end with the outer area of chamber 580.
• Rigid container 500 further comprises outlet channel 554 extending radially from core 600 to the more central portion of chamber 580 (i.e., the narrow portion or "neck” ofthe figure eight cross-section) and is fluidly connected at its distal end with chamber 580.
• inlet and outlet channels 550, 554 are shown in Figure 45 as being fluidly connected to the top end of chamber 580, the present invention also includes embodiments wherein both channels 550, 554 are in fluid communication with the bottom end of chamber 580, or wherein channel 550 is in fluid communication with the top end of chamber 580 and channel 554 is in fluid communication with the bottom end of chamber 580, or vice versa.
• Inlet and outlet channels 550, 554 are fluidly connected to dual lumen tubing 228 having an inlet lumen 230 and an outlet lumen 232.
• Rigid container 500 is removably secured to the upper surface 133 of upper bearing assembly 130 with appropriate screws, fasteners or the like (not shown).
• Inlet lumen 230 may be connected to a source for fluid medium, and outlet lumen 232 may be connected to a suction means for withdrawing the desired fraction from the chamber 580.
• chamber 580 is specifically designed to maximize the collection of platelet rich plasma by centrifugation of anticoagulated whole blood. More particularly, the shape of chamber 580 increases the width ofthe platelet rich plasma fraction when viewed from the top and decreases the depth of the platelet rich plasma fraction when viewed from the side, thus allowing the withdrawal of a greater amount of platelet rich plasma.
• Figure 44 shows a side profile of a rigid centrifuge container 500 as shown in Figure 43, having a generally oval profile and containing whole blood that has been separated into four fractions by centrifugation.
• width W 3 indicates the relative horizontal width ofthe platelet rich plasma fraction to be harvested
• D t indicates the relative depth ofthe platelet rich plasma fraction.
• Figure 47 shows a side profile of rigid centrifuge container 580 of this invention having the above-described off- centered figure eight shape and containing whole blood that has been separated into four fractions by centrifugation.
• width W indicates the relative horizontal width ofthe platelet rich plasma fraction 260 to be harvested
• D 2 indicates the relative depth ofthe platelet rich plasma fraction 260.
• Width W is necessarily wider than width W 3 in Figure 44.
• dumbbell shaped profile of chamber 580 shown in Figure 47 significantly increases the width W while decreasing the average depth D 2 , and therefore a greater portion ofthe platelet rich plasma fraction 260 can be withdrawn with greater accuracy before the platelet poor plasma fraction 262 reaches the outlet tube 554.
• chamber 580 In an embodiment where the platelet rich plasma is to be collected one could design chamber 580 as follows.
• the configuration of chamber 580 that is, the relative heights A, B, and C as shown in Figure 46, will be determined based on the typical location of the platelet rich plasma fraction 260 after centrifugation of whole blood.
• the platelet rich plasma will collect in chamber 580 at a region at a radial position ranging from about 35 to about 60 mm from the axis.
• the chamber 580 has a height of about 10 mm such that the horizontal surface area "B" of this region, illustrated in Figure 46, is about 4 mm 2 . Consequently, it can be appreciated that because ofthe unique configuration of chamber 580, the surface area of the platelet rich plasma fraction 260 as illustrated in Figure 47 may be maximized without undesirably increasing the overall size ofthe rigid centrifuge container 500. It will be appreciated by those skilled in the art that various geometric designs may be utilized depending on the fluid medium being centrifuged and the cellular fraction to be collected. The process for harvesting platelets from whole blood using rigid container 500 may be achieved in a manner similar to that described for bag 226
• Rigid centrifuge container 500 may be made from any number of rigid, transparent materials that are capable of withstanding typical sterilization conditions, including but not limited to acrylic resins, polycarbonate, or any clear thermal plastic.
• rigid container 500 is made of a cost-effective material that is relatively inexpensive to dispose of.
• centrifuge rotor having a complex interior geometry
• FIGs 48-52 An alternate embodiment of a centrifuge rotor of this invention for holding flexible centrifuge bag 226 is illustrated in Figures 48-52.
• the centrifuge rotor 755 is defined by a rotor base 760 ( Figures 48, 50 and 52) having a lower channel 780, and a rotor cover 770 ( Figures 49 and 51) having an upper channel 782.
• the annular interior chamber 784 ( Figure 48) of rotor 755 is defined by lower and upper channels 780, 782, and has a generally off-centered figure eight side cross-sectional configuration specifically designed to maximize the collection of platelet rich plasma by centrifugation of anticoagulated whole blood, as discussed below in detail.
• rotor base 760 comprises raised annular rim 775 and raised column 786 which is axially disposed in the interior of rotor base 760.
• Raised column 786 further has a groove 790 ( Figure 52) extending the diameter of column 786.
• the height of rim 775 is equal to the height of column 786.
• rotor cover 770 comprises raised annular rim 777 and raised column 788 which is axially disposed in the interior of cover 770.
• Raised column 788 further has a groove 792 ( Figure 52) extending the diameter of column 788.
• the height of rim 777 is equal to the height of column 788.
• Rotor base 760 and rotor cover 770 are preferably made from any number of rigid transparent materials including, but not limited to acrylic resins, polycarbonate, or any clear thermal plastic.
• centrifuge rotor 755 When centrifuge rotor 755 is to be assembled for use, flexible, doughnut-shaped centrifuge bag 226 having a center core 242 is placed in rotor base 760 such that center column 786 preferably, but not necessarily, extends through the core of centrifuge bag 226, and inlet and outlet tubes 248, 250 of bag 226 are seated in groove 790.
• rims 775 and 777 are in complete contact with each other, and columns 786 and 788 are preferably in complete contact with each other, thereby creating chamber 784 ( Figure 48).
• cover 770 is secured to base 760 as described, the inner perimeter of bag 226 is secured between columns 786 and 788 such that the columns do not physically contact each other.
• the generally flat, flexible centrifuge bag 226 When the generally flat, flexible centrifuge bag 226 is contained within chamber 784 prior to the infusion of a fluid medium (e.g., whole blood), it will not fill the entire volume of chamber 784 but rather will have a radially extending, flat shape as centrifuge rotor 755 is spinning. However, after a sufficient volume of the fluid medium (e.g., whole blood) has been introduced into flexible bag 226 through inlet tube 248 such that bag 226 is substantially completely filled, it will be appreciated that filled centrifuge bag 226 will conform to the shape' of chamber 784 and consequently will have a off-centered figure eight shaped cross-section.
• a fluid medium e.g., whole blood
• the off-centered figure eight configuration ofthe chamber 784 is of approximately the same configuration as the rigid bag 500. Therefore, for the same reasons, the shape of chamber 784 (and consequently the shape of filled bag 226), will assume an off-centered figure eight shape wherein the width of the platelet rich plasma fraction is greatly increased relative to the width of a filled bag having an elliptical cross-sectional shape
• the separation of platelet rich plasma fraction may be indicated by visual observation of a concentric ring containing the platelet rich plasma (which will be a less colored fraction) and an outer red-colored concentric ring containing the red blood cells.
• the platelet rich plasma may be withdrawn from centrifuge bag 226 by bent fitting 160 to direct the platelet rich plasma to the appropriate collector.
• sensors may be inco ⁇ orated as discussed in detail below to detect the presence ofthe platelet rich plasma fraction.
• the centrifugal processing system 10 and the centrifuge rotors and bags of this invention have a plurality of features which are suited to harvesting platelet rich plasma, white blood cells, platelet poor plasma or red blood cells from a patient's whole blood in accordance with each ofthe aspects of the present invention.
• hematocrits the volume of blood occupied by red blood cells, expressed as a percentage *
• the exact radial location of various blood components within the centrifuge bag after centrifugation will also vary.
• centrifuge bags of this invention overcome this issue by having an inlet tube capable of not only introducing whole blood into the centrifuge bag, but also capable of withdrawing some of all ofthe red blood cell fraction as needed to shift the location ofthe fraction to be harvested into the area of the outlet tube. Such features are presented in centrifuge bags 226, 270, 320 and 500. Yet another embodiment ofthe centrifuge bags of this invention which overcomes problems with varying hematocrits is centrifuge bag 226' having multiple outlet tubes.
• the centrifugal processing system 10 effectively provides a closed system which enhances the potential for maintaining a desired degree of sterility associated with the entire procedure since materials can thus be both provided to and removed from the centrifuge bag during rotation ofthe centrifuge via, for instance, a dual lumen tubing connected to a fluid source (e.g., anticoagulated whole blood withdrawn from a patient before or during surgery) and collection containers (i.e., for the preparation of a platelet gel), without interrupting the process, and thus without significant exposure of the materials to environmental conditions.
• a fluid source e.g., anticoagulated whole blood withdrawn from a patient before or during surgery
• collection containers i.e., for the preparation of a platelet gel
• the portable size ofthe centrifugal processing system 10 in combination with the above-described features of shifting the separated fractions and maximizing the surface area ofthe harvested fraction allows for increased processing capabilities autologous platelet gel over larger, conventional centrifuges
• the on-line harvesting capabilities ofthe centrifugal processing system 10 allows for continuous, dynamic separation and collection of platelet rich plasma, white blood cells, red blood cells and platelet poor plasma, by adjusting the input and removal of fluid medium and separated fractions as described above. Further, the orientation of the flexible and rigid centrifuge bags of this invention and ofthe contents therein (e.g., being generally radially extending) is not significantly modified in the transformation from separation to harvesting of the various constituents. Moreover, vortexing throughout the contents of the centrifuge bags of this invention is reduced or eliminated since the centrifugal processing system 10 does not have to be decelerated or stopped for addition of fluid medium or removal ofthe various fractions therefrom.
• the general orientation ofthe flexible and rigid centrifuge bags ofthe invention (e.g., substantially horizontal) is maintained during removal of the desired whole blood fraction similar to the orientation ofthe centrifuge bags assumed during centrifugation to further assist in maintaining the degree of separation provided by centrifugation. Consequently, the potential is reduced for disturbing the fractions to the degree where the separation achieved is adversely affected.
• a further embodiment of the present invention provides for the automation of at least portions of the separation and material handling processes.
• an automated centrifugal processing system 800 is illustrated that is generally configured to provide automated control over the steps of inputting blood, separating desired components, and outputting the separated components.
• the following discussion ofthe processing system 800 provides examples of separating platelets in a blood sample, but the processing system 800 provides features that would be useful for separating other components or fractions from blood or other fluids. These other uses for the processing system 800 are considered within the breadth of this disclosure.
• the specific components discussed for use in the processing system 800 are provided for illustration pu ⁇ oses and not as limitations, with alternative devices being readily apparent to those skilled in the medical device arts.
• the processing system 800 includes a blood source 802 connected with a fluid line 804 to an inlet pump 810.
• the inlet pump 810 is operable to pump blood from the blood source 802 through the fluid line 818 to a centrifuge 820. Once all or a select portion of the blood in the blood source 802 have been pumped to a blood reservoir 824 of the centrifuge 820 the inlet pump 810 is turned off and the blood source 802 isolated with valve 806.
• the inlet pump 810 may be operated at later times to provide additional blood during the operation ofthe processing system 800 (such as during or after the removal of a separated component).
• the centrifuge 20 preferably includes a flexible centrifuge bag, for example 226, 226', 270, or 320, positioned within the rotor 202 for collecting the input blood, or alternatively rotor 202 may be a rigid container having an off centered figure eight shaped chamber, which may collect blood directly as discussed previously.
• a centrifuge bag 226 it is to be understood that the alternative centrifuge bags disclosed herein may be used in a similar manner.
• the centrifuge 20 as discussed above has an internal mid-shaft gear assembly 108 that provides the motive force to rotate the rotor assembly 200, and particularly the rotor 202, at a rotation rate that is adequate to create centrifugal forces that act to separate the various constituents or components of the blood in the rotor 202.
• the drive assembly 822 may comprise a number of devices useful for generating the motive force, such as an electric motor with a drive shaft connected to internal drive components ofthe centrifuge 20.
• the drive assembly 822 comprises an electric motor that drives a belt attached to an exterior portion ofthe centrifuge 20 and more particularly to the timing belt ring 44. To obtain adequate separation, the rotation rate is typically between about 0
• RPM and 5000 RPM is maintained between about 0 RPM and 5000 RPM.
• components of particular densities assume radial positions or belts at differing distances from the central axis A ofthe rotor 202.
• the heavier red blood cells typically separate in an outer region while lower density platelets separate into a region more proximal to the central axis of the rotor 202.
• this density interface is used in some embodiments ofthe centrifugal processing system 10 to identify the location of component regions (as will be discussed in more detail below).
• the drive assembly 822 continues to operate to rotate the centrifuge 20 to retain the separation of the components throughout the operation ofthe centrifugal processing system 10.
• the outlet pump 830 is operated to pump select components from the rotor 202 through outlet lumen 828.
• the centrifuge bag held within the rotor 202 preferably is configured to allow the selective removal of a separated blood component, such as platelets located in a platelet rich plasma region, by the positioning of an outlet lumen 232 a radial distance from the central axis of the centrifuge bag 226.
• this radial distance or radial location for the outlet lumen is selected to coincide with the radial location ofthe desired, separated component or the anticipated location of the separated component.
• the outlet pump 830 only (or substantially only) removes a particular component (such as platelets into container 400) existing at that radial distance. Once all or a desired quantity ofthe particular component is removed from the centrifiige bag 226, operation ofthe outlet pump 830 is stopped, and a new separation process can be initiated. Alternatively, in a preferred embodiment, additional blood is pumped into the centrifuge by 226 by further operating the inlet pump 810 after or concurrent with operation ofthe outlet pump 830.
• each blood sample may have varying levels or quantities of different components.
• the radial distance or location of a particular component or component region within the centrifuge bag 226 varies, at least slightly, with each different blood sample.
• the size of the component region also varies and the amount that can be pumped out of the centrifuge bag 226 by the outlet pump 830 without inclusion of other components varies with each blood sample.
• the position of the component region will vary in embodiments of the separation system 10 in which additional blood is added after or during the removal of blood by the outlet pump 830.
• the centrifugal processing system 10 preferably is configured to adjust the location of a separated component to substantially align the radial location of the separated component with the radial location of the outlet port.
• the centrifugal processing system 10 may be utilized to collect platelets from a blood sample.
• the centrifugal processing system 10 preferably includes a red blood cell collector 812 connected to the inlet pump 810 via fluid line 814 having an isolation valve 816 (e.g., a solenoid-operated valve or one-way check valve).
• the pump or syringe may also act as the valve.
• the inlet pump 810 is configured to selectively pump fluids in two directions, to and away from the centrifuge 820 through fluid line 818, and in this regard, may be a reversible-direction peristaltic pump or other two-directional pump.
• a single fluid line may be utilized as an inlet and an outlet line to practice the invention. Operation ofthe inlet pump 810 to remove fluid from the centrifuge bag 226 is useful to align the radial location ofthe desired separated component with the outlet tube 250 and inlet tube 248 ofthe centrifuge bag 226.
• inlet tube 248 connected to lumen 232 and 230, respectively, inlet tube 248 is preferably at a greater radial distance than the outlet tube 250.
• suction is applied to the inlet lumen 230 by inlet pump 810, red blood cells are pumped out ofthe centrifuge bag 226 and into the red blood cell collector 812. As red blood cells are removed, the separated platelets (i.e., the desired component region) move radially outward to a new location within the centrifuge bag 226.
• the inlet pump 810 is operated until the radial distance ofthe separated platelets or platelet region from the central axis is increased to coincide with the radial distance or location of the outlet tube 250 ofthe centrifuge bag 226. Once substantial alignment ofthe desired component region and the outlet tube 250 is achieved, the outlet pump 830 is operated to remove all or a select quantity ofthe components in the aligned component region.
• the centrifugal processing system 10 includes a controller 850 for monitoring and controlling operation ofthe inlet pump
• the controller 850 comprises a computer with a central processing unit (CPU) with a digital signal processor, memory, an input/output (I/O) interface for receiving input and feedback signals and for transmitting control signals, and software or programming applications for processing input signals and generating control signals (with or without signal conditioners and/or amplifiers).
• CPU central processing unit
• I/O input/output
• software or programming applications for processing input signals and generating control signals (with or without signal conditioners and/or amplifiers).
• the controller 850 is communicatively linked to the devices ofthe centrifugal processing system 10 with signal lines 860, 862, 864, 866, and 868 which may include signal conditioning devices and other devices to provide for proper communications between the controller 850 and the components ofthe centrifugal processing system 10.
• signal lines 860, 862, 864, 866, and 868 may include signal conditioning devices and other devices to provide for proper communications between the controller 850 and the components ofthe centrifugal processing system 10.
• the controller 850 After initiation ofthe centrifuge spinning or concurrently with operation ofthe drive assembly 822, the controller 850 generates a control signal over signal line 860 to the inlet pump 810 to begin pumping blood from the blood source container 802 to the centrifiige bag 226 ofthe centrifiige 20.
• the drive assembly 822 is operable at more than one speed or over a range of speeds. Additionally, even with a single speed drive shaft the rotation rate achieved at the centrifuge 20 may vary.
• the processing system 10 may include a velocity detector 858 that at least periodically detects movement ofthe centrifuge bag 226 portion ofthe centrifuge 20 and transmits a feedback signal over signal line 866 to the controller 850.
• the controller 850 processes the received signal to calculate the rotation rate of the centrifuge 20, and if applicable, transmits a control signal to the drive assembly 822 to increase or decrease its operating speed to obtain a desired rotation rate at the centrifuge bag 226.
• the processing system 800 may be calibrated to account for variations in the centrifuge 20 and drive assembly 822 configuration to determine a minimum rotation time to obtain a desired level of component separation.
• the controller 850 preferably includes a timer mechanism 856 that operates to measure the period of time that the centrifuge 20 has been rotated by the drive assembly 822 (such as by beginning measuring from the transmission of the control signal by the controller 850 to the drive assembly 822).
• the timing mechanism 856 informs the controller 850 that separation has been achieved in the centrifuge bag 226.
• the controller 850 operates to transmit control signal over signal line 860 to the input pump 810 to cease operation and to the outlet pump 830 over signal line 868 to initiate operation to pump a separated component in the component region adjacent the outlet port of lumen 232 of centrifuge bag 226 through fluid line 828.
• the velocity feedback signal from the velocity detector 858 is utilized by the controller 850 to adjust the rotation time as necessary to obtain the desired level of component separation.
• the centrifugal processing system 10 can be calibrated for a number of rotation rates and the corresponding minimum rotation times can be stored in a look up table for retrieval by the controller 850 based on a calculated rotation rate. Rotational rates may be varied either manually or automatically to optimize cellular component position and or concentration.
• a preferred embodiment of the centrifugal processing system 800 includes a sensor assembly 840 to monitor the separation of components within the centrifuge bag and to transmit feedback signals over line 862 to the controller 850.
• numerous sensor devices exist for detecting the presence of certain components in a fluid, and specifically a blood, sample. Many of these devices comprise a source of radiant energy, such as infrared, laser, or incandescent light, and a compatible radiant energy-sensitive detector that reacts to the received energy by generating an electric signal. Briefly, these radiant energy devices are useful because the detected signal varies in a measurable fashion with variances in the density ofthe material through which beams ofthe radiant energy are passed.
• the sensor assembly 840 may comprise any of these well-known types of radiant energy source and detector devices and other sensor devices useful for measuring the existence of constituents of fluids such as blood.
• the source and the detector ofthe sensor assembly 840 are preferably located within the centrifugal processing system 800 to allow monitoring of the centrifuge bag 226 and, particularly, to identify the presence of a particular blood component in a radial position coinciding with the radial position ofthe outlet port of the centrifuge bag 226.
• the radiation beams from the source are transmitted through a "window" in the centrifuge bag 226 that has a radial location that at least partially overlaps the radial location of the outlet port.
• the feedback signals from the detector of the sensor assembly 840 allow the controller 850 to identify when a density interface has entered the window. This may occur for a number of reasons.
• red blood cells When red blood cells are being removed by operation ofthe inlet pump 810 to remove fluid from the centrifuge bag 226 via the inlet tube 248.
• the change in density may also occur when a denser component is being added to the centrifuge bag 226 causing the particular blood component to be pushed radially inward. In the centrifugation of whole blood, this occurs when additional blood is added by operation ofthe input pump 810 and red blood cells collect in a region radially outward from the platelet region.
• the window of the radiation source may be alternatively positioned radially inward from the location ofthe outlet tube 250 ofthe centrifuge bag 226.
• the controller 850 can identify when the outlet pump 830 has nearly removed all of the particular component ofthe monitored region and/or when the inlet pump 810 has removed a quantity of denser components causing the monitored region to move radially outward.
• the controller 850 can then operate to send control signals to turn off the outlet pump 830 or the inlet pump 810 (as appropriate) to minimize the amount of undesired components (lower density components) that enter the outlet tube 250.
• the sensor assembly 840 may have two radiation sources and detectors, and the second window ofthe sensor assembly 840 may be located a distance radially outward from the outlet tube 250. With two sensing windows, the sensor assembly 840 is operable to provide the controller 850 information about a density interface moving radially inward toward the outlet tube 250 (such as when red blood cells are added). In response, the controller 850 can generate a control signal to the inlet pump 810 to operate to pump the denser components, such as red blood cells, out ofthe centrifuge bag 226.
• Two sensing windows also allow the controller 850 to detect a density interface moving outward, which allows the controller 850 to shut off the outlet pump 830 (and/or the inlet pump 810 to stop evacuating processes) and/or to start the inlet pump 810 to add additional blood.
• Figure 54 is provided which illustrates the timing and relationship of control signals generated by the controller 850 and the receipt of feedback signals from the sensor assembly 840.
• the radiation detector ofthe sensor assembly 840 is positioned adjacent outlet tube (inlet to the outlet pump 830) in the centrifuge bag 226 to sense density changes in the fluid flowing past the outlet tube 250.
• operation of the processing system 800 begins at time to, with the inlet pump 810, the outlet pump 830, and the centrifuge drive assembly 822 all being off or not operating.
• the controller 850 operates in response to operator input or upon sensing the blood source 802 is adequately filled (sensor not shown) to generate a control signal on line 864 to begin operating the centrifuge drive assembly 822 to rotate the centrifuge bag 226.
• this control signal over line 864 also contains rotation rate information to initially set the operating speed ofthe drive assembly 822.
• the controller 850 Concu ⁇ ently or at a selected delay time, the controller 850 generates a control signal on line 860 to start the inlet pump 810 in a configuration to pump fluid to the centrifuge bag 226 over fluid line 818.
• the sensor assembly 840 provides an initial density feedback signal to the controller 850 on line 862, which the controller 850 can process to determine an initial or unseparated density adjacent the outlet tube.
• the controller 850 may be configured to request a feedback signal from the sensor assembly 840 after a set delay period (as measured by the timer mechanism 856) to allow separation ofthe components being pumped into the centrifuge bag 226 (such as the calibrated, minimum rotation time discussed above) into regions.
• the controller 850 functions to align the region having the desired density, such as a region comprising a higher density of platelets, adjacent the detector of the sensor assembly 840 (i.e., adjacent the outlet tube). To achieve alignment, the controller 850 transmits a control signal over line 860 to the inlet pump 810 to stop pumping fluid to the centrifuge bag 226, to reverse pumping directions including shutting valve 806 and opening valve 816, and to begin pumping components having a higher density then the particular, desired component from the centrifuge bag 226 to the collector 812.
• the region having the desired density such as a region comprising a higher density of platelets
• the inlet pump 810 when the centrifugal processing system 10 is operated to separate and collect platelets or platelet rich plasma, the inlet pump 810 at time, t 2 , is operated to pump out the red blood cell fraction by applying suction at the inlet tube 248 to the centrifuge bag 226.
• the density ofthe fluid adjacent the outlet tube 250 begins to change as denser components are removed by the inlet pump 810, and the sensor feedback signal being transmitted to the controller 850 changes in magnitude.
• the sensor feedback signal continues to change in magnitude (either becoming stronger or weaker depending on the particular sensor utilized and the material being collected) until at time , when the controller 850 processes the feedback signal and determines that the density of the adjacent fluids is within a desired range. This transition can also be thought of as detecting when an interface between two regions of differing densities passes by the location ofthe detector of the sensor assembly 840.
• the controller 850 operates at time , to send a control signal over line 860 to stop operations of the inlet pump 810. Also, at time , or at any time thereafter, the controller 850 generates a control signal over line 868 to begin operation the outlet pump 830 to apply suction at the outlet tube 250 ofthe centrifuge bag 226 to remove the desired component, such as the platelet rich plasma fraction, from the centrifuge bag 226.
• the sensor feedback signal again begins to change in magnitude as the density ofthe fluid near the outlet tube 250 begins to change, such as when platelet poor plasma begins to enter the sampling window ofthe sensor assembly 840.
• the controller 850 transmits a control signal on line 868 to halt operations ofthe outlet pump 830.
• the controller 850 can be operated to transmit the signal to the outlet pump 830 at any time prior to time t ⁇ , such as at a time after time ts, when the density of the adjacent fluid begins to change but prior to time te or based on volume removed.
• the controller 850 can then operate any time after time , to halt operation ofthe centrifuge drive assembly 822.
• operations ofthe separation centrifugal processing system 800 can be repeated with the inlet pump 810 being operated to add additional fluid, e.g., blood, after time t 6 .
• the inlet pump 810 and the outlet pump 830 may be operated concurrently to add an additional volume of blood with a corresponding new amount of the component being collected after time tt, to extend the period of time between detection of the interface at time U and the detection of an out of range density at time t 6 .
• a sensor assembly 840 was shown in Figure 53 schematically, and it was noted that the location of a radiant energy source and a detector may be any location within the processing system 800 useful for obtaining an accurate measurement of separating blood components within the centrifuge bag 226.
• the source and detector can be both positioned within the centrifuge 20 at a location adjacent the centrifuge bag 226.
• problems may arise with providing proper signal and power line connections to the source and sensor and with accounting for the rotation of the centrifuge and portions ofthe sensor assembly 840.
• one preferred embodiment of the processing system 800 provides for an externally positioned sensor assembly 840 including source and detector to simplify the structure ofthe centrifuge 20 while still providing effective density determinations of fluids within the blood reservoir.
• FIG 55 illustrates a general side view of the relevant components of this external sensor embodiment of the centrifugal processing system 800.
• the centrifuge 20 comprises a rotor extension portion 880 and a drive portion 881, which is connected to the drive assembly 822 (connection not shown). Both the centrifuge 20 and the rotor extension portion 880 rotate about a central or rotation axis, C a ⁇ ls , of the centrifuge 20.
• the drive portion 881 spins in a ratio of 2 to 1 (or other suitable ratio) relative to the reservoir extension portion 880 to control twisting of inlet and outlet fluid lines to the rotor extension portion 880.
• the internal gearing features of the centrifuge 20 also enable the centrifuge 20 to effectively obtain rotation rates that force the separation of components with differing densities while limiting the risk that denser components, such as red blood cells, will become too tightly packed during separation forming a solid, dense material that is more difficult to pump or remove from the centrifuge 20.
• the rotor extension portion 880 is shown located on the upper end of the centrifuge 20 and includes a centrifuge bag 226 or other receptacle.
• the rotor extension portion 880 is fabricated from a transparent or partially transparent material, such as any of a number of plastics, to allow sensing of fluid densities.
• the rotor extension portion 880 extends a distance, d ove r, beyond the outer edge of the centrifuge 20 as measured radially outward from the central axis, Ca ⁇ , s .
• the distance, d 0Ver is preferably selected such that the desired component, such as the platelet rich plasma fraction, to be collected readily separates into a region at a point within the centrifuge bag 226 that also extends outward from the centrifuge 20.
• the rotor extension portion 880 is also configured so that the centrifuge bag 226 extends within the rotor extension portion 880 to a point near the outer circumference of the rotor extension portion 880.
• the distance, dov er selected for extending the rotor extension portion 880 is preferably selected to facilitate alignment process (discussed above) and to control the need for operating the input pump 810 to remove denser components.
• the distance, d ove r is selected such that during separation of a typical blood sample center ofthe platelet rich region is about one half the extension distance, d over , from the circumferential edge ofthe centrifuge 20.
• the sensor assembly 840 is entirely external to the centrifuge 20 as shown in Figure 55.
• the sensor assembly 840 includes a source 882 for emitting beams 884 of radiant energy into and through the rotor extension portion 880 and the included centrifuge bag 226.
• the radiant energy source 882 may be nearly any source of radiant energy (such as incandescent light, a strobe light, an infrared light, laser and the like) useful in a fluid density sensor and the particular type of detector or energy used is not as important as the external location ofthe source 882.
• the sensor assembly 840 further includes a detector 886 that receives or senses beams 888 that have passed through the centrifuge bag 226 and have impinged upon the detector 886.
• the detector 886 receives or senses beams 888 that have passed through the centrifuge bag 226 and have impinged upon the detector 886.
• the detector 886 is selected to be compatible with the source 882 and to transmit a feedback signal in response sensing the energy beams 888.
• the detector 886 (in combination with the controller 850 and its processing capacities) is useful for detecting the density of fluids in the centrifuge bag 226 between the source 882 and the detector 886.
• the sensor assembly 840 is useful for identifying changes in fluid density and interfaces between fluids with differing densities. For example, the interface between a region containing separated red blood cells and a region containing the platelet rich plasma fraction, and the interface between the platelet rich plasma region and a platelet-poor plasma region.
• a sampling window is created rather than a single sampling point (although a single sampling point configuration is useful as part ofthe invention as creating a window defined by a single radial distance).
• the sampling window is defined by an outer radial distance, dou ⁇ , from the central axis, Ca ⁇ ls and an inner radial distance, d ⁇ ».
• the size ofthe sampling window may be rather small approximating a point and may, of course vary in cross-sectional shape (e.g., circular, square, rectangular, and the like).
• the sensor assembly 840 be positioned relative to the reservoir extension portion 880 and the centrifuge bag 226 such that the sampling window created by the source 882 and detector 886 at least partially overlaps the radial position ofthe region created during separation processes containing a component of particular density, such as platelets.
• This may be a calibrated position determined through calibration processes ofthe centrifuge 20 in which a number of blood (or other fluid) samples are fully separated and radial distances to a particular region are measured. The determined or calibrated position can then be utilized as a initial, fixed location for the sensor assembly 840 with the source 882 and detector 886 being positioned relative to the rotor extension portion 880 such that the sampling window overlaps the anticipated position ofthe selected separation region.
• each sample may vary in content of various components which may cause this initial alignment to be inaccurate and operations ofthe centrifugal processing system 800 may cause misalignment or movement of regions.
• alignment processes discussed above preferably are utilized in addition to the initial positioning ofthe sampling window created by the sensor assembly 840.
• the sensor assembly 840 is not in a fixed position within the separation system 800 and can be positioned during separation operations.
• the sensor assembly 840 may be mounted on a base which can be slid radially inward toward the centrifuge 20 and radially outward away from the centrifuge 20 to vary the distances, di and dou ⁇ - This sliding movement is useful for providing access to the centrifuge bag 226, such as to insert and remove a disposable bag.
• the sensor assembly 840 would initially be pushed outward from the centrifuge 20 until a new bag was inserted into the centrifuge bag 226. The sensor assembly 840 could then be slid inward (or otherwise moved inward) to a calibrated position.
• the centrifugal processing system 800 could be operated for a period of time to achieve partial or full separation (based on a timed period or simple visual observation) and then the sensor assembly 840 slid inward to a position that the operator ofthe centrifugal processing system 800 visually approximates as aligning the sampling window with a desired region of separated components (such as the platelet rich plasma region). The effectiveness of such alignment could then readily be verified by operating the sensor assembly 840 to detect the density of the fluids in the centrifuge bag 226 and a calculated density (or other information) could be output or displayed by the controller 850.
• This alternate embodiment provides a readily maintainable centrifugal processing system 800 while providing the benefits of a fixed position sensor assembly 840 and added benefits of allowing easy relative positioning to obtain or at least approximate a desired sample window and separation region alignment.
• rotor extension portion 880 it may be preferable to not have a rotor extension portion 880 or to modify the rotor extension portion 880 and the sensor assembly 840 such that the extension is not significant to monitoring the separation within the blood reservoir or centrifuge bag 226.
• Two alternative embodiments or arrangements are illustrated in
• FIGS 56 and 57 that provide the advantages of an external sensor assembly 840 (such as an external radiation source and detector). With these further embodiments provided, numerous other expansions of the discussed use of an external sensor will become apparent to those skilled in the arts and are considered within the breadth of this invention.
• a rotor 202 is illustrated that has no extending portion
• the rotor 202 and centrifuge bag 226 are preferably fabricated from plastics or other materials that allow radiation to pass through to detect changes in densities or other properties of fluid samples within the centrifuge bag 226.
• the radiation source 882 and the detector 886 are not positioned on opposing sides ofthe rotor 202. Instead, a reflector 885 (such as a mirror and the like) is positioned within the drive portion 881 of the centrifuge to receive the radiation beams 884 from the radiation source 882 and direct them through the portion 880 and centrifuge bag 226.
• the detector 886 is positioned within the sensor assembly 840 and relative to the centrifuge 20 to receive the deflected or reflected beams 888 that have passed through the fluid sample in the centrifuge bag 226.
• the sampling window within the centrifuge bag 226 can be selected to align with the anticipated location ofthe fraction that is to be collected upon separation.
• the sampling window at least partially overlaps with the location of the outlet tube ofthe blood reservoir or centrifuge bag 226.
• the drive portion is fabricated from a non-transparent material and a path for the beams 884 from the radiation source 884 to the reflector 885 is provided.
• the path in one prefe ⁇ ed embodiment is an opening or hole such as port 154 or 156 ( Figure 14) in the side of the drive portion 881 that creates a path or tunnel through which the beams 884 travel unimpeded.
• the opening may be replaced with a path of transparent material to allow the beams to travel to the reflector 885 while also providing a protective cover for the internals of the drive portion 881.
• a path is also provided downstream ofthe reflector 885 to allow the beams 884 to travel through the drive portion 881 internals without or with minimal degradation.
• the path may be an opening or tunnel through the drive portion leading to the portion 202 or be a path created with transparent materials.
• the beams 884 in these tunnel path embodiments enter the drive portion 881 one time per revolution of the drive portion 881, which provides an acceptable rate of sampling.
• a reflector 885 may readily be provided that extends circumferentially about the center axis of the drive portion 881 to provide a sampling rate equivalent to the rate of beam 884 transmission.
• the positions of the radiation source 882 and the detector 886 may be reversed and the angle ofthe reflector 885 and transmission ofthe beams 884 may be altered from those shown to practice the invention.
• FIG. 57 A further embodiment of an external sensor assembly 840 is provided in Figure 57.
• the radiation source 882 also acts as a radiation detector so there is no need for a separate detector.
• the radiation source and detector 882 transmits beams 884 into the rotating drive portion 881 through or over the path in the drive portion 881.
• the reflector 885 reflects the beams 884 toward the rotor 202 and the centrifuge bag 226 to create a sampling window within the centrifuge bag 226 in which density changes may be monitored.
• the beams 888 strike a second reflector 887 that is positioned within the rotor 202 to reflect the beams 888 back over the same or substantially the same path through the centrifuge bag 226 to again strike the reflector 885.
• the reflector 885 directs the beams 888 out of the drive portion 881 and back to the radiation source and detector 882 which, in response to the impinging beams 888, transmits a feedback signal to the controller 850 for further processing.
• the beams 884 enter the driving portion 881 once during every revolution ofthe driving portion 881.
• the portion 880 is preferably rotating twice for every rotation of the driving portion 881, as discussed in detail above, and hence, the second reflector 887 is aligned to receive the beams 888 only on every other rotation of the driving portion 881.
• a pair of reflectors 887 may be positioned in the rotor 202 such that the beams 888 may be received and reflected back through the centrifuge bag 226 once for every rotation ofthe driving portion 881.
• the reflector 885 and second reflector 887 may expand partially or fully about the center axis ofthe centrifuge 20 (with co ⁇ esponding openings and/or transparent paths in the driving portion 881) to provide a higher sampling rate.
• temperature control features are provided in an alternate embodiment ofthe automated processing system invention 900, as illustrated in Figure 58.
• Providing temperature controls within the processing system 900 can take many forms such as controlling the temperature of input fluid samples from the blood source 802, monitoring and controlling the temperature of fluids in the centrifuge bag 226 to facilitate separation processes, and controlling the operating temperature of temperature sensitive components of the processing system 900. These components include but are not limited to, red blood cells, white blood cells, plasma, platelet rich plasma or any of these components mixed with other drugs, proteins or compounds.
• a temperature control system is included in the processing system 900 to heat components removed from the centrifuge bag 226 by the outlet pump 830 to a desired temperature range.
• a dispenser assembly 902 is included in the processing system 900 and includes chambers or syringes for collecting and processing platelet rich plasma drawn from the centrifuge 20.
• the temperature control system is useful in this regard for raising, and for then maintaining, the temperature ofthe platelets in the dispenser assembly to a predetermined activation temperature range.
• the activation temperature range is 25° C to 50° C and preferably 37° C to 40° C, but it will be understood that differing temperature ranges may readily be utilized to practice the invention depending on the desired activation levels and particular products being processed or created with the processing system 900.
• the temperature control system ofthe processing system 900 includes a temperature controller 904 that is communicatively linked to the controller
• the controller 850 may be utilized to initially set operating temperature ranges (e.g., an activation temperature range) and communicate these settings over feedback signal line 906 to the temperature controller 904.
• the temperature controller 904 may include input/output (I/O) devices for accepting the operating temperature ranges from an operator or these ranges may be preset as part ofthe initial fabrication and assembly ofthe processing system 900.
• the temperature controller 904 may comprise an electronic control circuit allowing linear, proportional, or other control over temperatures and heater elements and the like.
• the temperature controller 904 includes a microprocessor for calculating sensed temperatures, memory for storing temperature and control algorithms and programs, and I/O portions for receiving feedback signals from thermo sensors and for generating and transmitting control signals to various temperature control devices (e.g., resistive heat elements, fan rotors, and other devices well-known to those skilled in the heating and cooling arts).
• a temperature sensor 908 comprising one or more temperature sensing elements is provided to sense the temperature ofthe dispenser assembly 902 and to provide a co ⁇ esponding temperature feedback signal to the temperature controller 904 over signal line 910 (such as an electric signal proportional to sensed temperature changes).
• the temperature sensor 908 may be any temperature sensitive device useful for sensing temperature and, in response, generating a feedback signal useful by the temperature controller 904, such as a thermistor, thermocouple, and the like.
• the temperature sensor 908 is positioned within the dispenser assembly 902 to be in heat transferring or heat sensing contact with the syringes or other chambers containing the separated product which is to be activated. In this manner, the temperature controller 904 is able to better monitor whether the temperature ofthe relevant chambers within the dispenser assembly 902 is within the desired activation temperature range.
• a heater element 913 is included in the temperature control system and is selectively operable by the temperature controller 904 such as by operation of a power source based on signals received from the temperature sensor 908.
• the heater element 913 may comprise any number of devices useful for heating an object such as the chambers ofthe dispenser assembly 902, such as a fluid heat exchanger with tubing in heat exchange contact with the chambers.
• electrical resistance-type heaters comprising coils, plates, and the like are utilized as part ofthe heater element 913.
• the resistive portions ofthe heater element 913 would be formed into a shape that conforms to the shape of the exterior portion of the chambers of the dispenser assembly 902 to provide efficient heat transfer but preferably also allow for insertion and removal of the chambers ofthe dispenser assembly 902.
• the temperature controller controls the temperature of the heater element 913 to provide efficient heat transfer but preferably also allow for insertion and removal of the chambers ofthe dispenser assembly 902.
• the 904 is configured to receive an operating temperature range, to receive and process temperature feedback signals from the temperature sensor 908, and in response, to selectively operate the heater element 913 to first raise the temperature ofthe chambers of the dispenser assembly 902 to a temperature within the operating temperature range and to second maintain the sensed temperature within the operating range.
• a desired operating range for activating a gel or manipulating other cellular components and their reactions onto themselves or with agents may be provided as a set point temperature (or desired activation temperature) with a tolerance provided on either side of this set point temperature.
• the temperature controller 904 may operate the heater element 913 to raise the temperature ofthe chambers of the dispenser assembly 902 to a temperature above the set point temperature but below the upper tolerance temperature at which point the heater element 913 may be shut off by the temperature controller 904.
• the temperature controller 904 operates the heater element 913 to again raise the sensed temperature to above the set point temperature but below the upper tolerance temperature.
• the temperature controller 904 effectively maintains the temperature of the chambers in the dispenser assembly 902 within a desired activation temperature range (which, of course, may be a very small range that approximates a single set temperature).
• the temperature controller is or operates as a proportional integral derivative (PID) temperature controller to provide enhanced temperature control with smaller peaks and abrupt changes in the temperature produced by the heater element 913.
• the temperature controller 904 may include visual indicators (such as LEDs) to indicate when the sensed temperature is within a set operating range and/or audio alarms to indicate when the sensed temperature is outside the set operating range.
• the heater element 913 is configured to operate at more than one setting such that it may be operated throughout operation of the processing system 900 and is not shut off.
• the heater element 913 may have a lower setting designed to maintain the chambers of the dispenser assembly 902 at the lower end of the operating range (e.g., acceptable activation temperature range) with higher settings that provide heating that brings the chambers up to higher temperatures within the set operating range.
• the heater element 913 is configured to heat up at selectable rates (e.g., change in temperature per unit of time) to enhance the activation or other processing of separated liquids in the dispenser assembly 902. This feature provides the temperature controller 904 with control over the heating rate provided by the heater element 913.
• Figure 59 illustrates one preferred arrangement ofthe centrifugal processing system 900 of Figure 58 that provides a compact profile or footprint while facilitating the inclusion of a temperature control system.
• An enclosure 916 is included as part ofthe temperature control system to provide structural support and protection for the components ofthe temperature control system.
• the enclosure 916 may be fabricated from a number of structural materials, such as plastic.
• the enclosure 916 supports a heater housing 918 that is configured to allow insertion and removal ofthe chambers and other elements of the dispenser assembly 902.
• the heater housing 918 has a wall that contains the heater element 913 (not shown in Figure 59) which is connected via control line 914 to the temperature controller 904.
• the temperature sensor 908 (not shown in Figure 59) is also positioned within the heater housing 918, and as discussed with reference to Figure 58, is positioned relative to the chambers ofthe dispenser assembly 902 to sense the temperature of the chambers, and the contained fluid, during operation ofthe system 900.
• a temperature feedback signal is transmitted by the temperature sensor 908 over line 910 to temperature controller 904, which responds by selectively operating the heater element 913 to maintain the temperature within the heater housing 918 within a selected operating range.
• the temperature control system preferably is configured to monitor and control the temperature within the enclosure 916.
• a temperature sensor 920 is included to sense the ambient temperature within the enclosure 916 and to transmit a feedback signal over line 922 to temperature controller 904.
• An air inlet 930 such as a louver, is provided in the enclosure 916 to allow air, A IN , to be drawn into and through the enclosure 916 to remove heated air and maintain the temperature within the enclosure 916 at an acceptable ambient temperature.
• a fan 934 is provided to pull the air, A IN , into the enclosure 916 and to discharge hotter air, A OUT , out ofthe enclosure 916.
• the fan 934 is selectively operable by the temperature controller 904 via control signals over line 938.
• the size or rating ofthe fan 934 may vary in embodiments of the invention and is preferably selected based on the volume of the enclosure 916, the components positioned within the enclosure 916 (e.g., the quantity of heat generated by the separation system 900 components), the desired ambient temperature for the enclosure 916, and other cooling design factors.
• a dispenser 902 for manipulating the cellular fraction which has been isolated and collected via outlet lumen 232.
• the present invention relates to a dispenser 902 which allows for a manual or automated manipulation of a two-phase method for forming an autologous platelet gel 970 composition wherein all ofthe blood components for the platelet gel 970 are derived from a patient to whom the platelet gel 970 will be applied.
• the methods ofthe present invention for preparing an autologous platelet gel 970 composition discussed in further detail below, are represented in the flow diagrams depicted in Figures 61-63.
• the methods of the present invention begins by forming anticoagulated whole blood 396 which is achieved by collecting a patient's whole blood 394 in a source container 398 having an anticoagulation agent, such as sodium citrate ( mecanicte) or heparin.
• an anticoagulation agent such as sodium citrate (gulte) or heparin.
• the whole blood 394 is collected and mixed with a 3.8% solution of sodium citrate (referred to herein as "citrate collection medium") specifically in a 9:1 ratio of blood to citrate collection medium.
• a 3.8% solution of sodium gulte is prepared by adding 3.8 grams of sodium gulte per 100 ml of water. While a 3.8% sodium citrate collection medium is that which is frequently used to collect and preserve blood, the person skilled in this art will recognize that the ratio of sodium citrate to whole blood could be in the range of about 10.9 - 12.9% mMOL, final concentration.
• platelet rich plasma 260 and/or platelet poor plasma 262 are formed by centrifuging a quantity of anticoagulated whole blood 396 that was previously drawn from the patient.
• the platelet rich plasma 260 is first drawn from the centrifuge bag 226 and into collection chamber
• Collection chamber 400 is preferably a syringe, but any container that will not contact activate the collected fraction is acceptable.
• the platelet rich plasma 260 can be pumped via outlet pump 830 ( Figure 53) into a collection chamber 400 or the desired fraction can be drawn directly into dispenser 902.
• the platelet rich plasma 260 in centrifuge bag 226, is divided into two portions and stored in vessels 952 and 960. The first portion is approximately l A to Vz ofthe total volume of platelet rich plasma 260 and is utilized in phase-one to prepare the thrombin, while the second portion of platelet rich plasma 260 is utilized in phase-two vessel 960.
• the prefe ⁇ ed methods to obtain thrombin and then produce the platelet gel compositions in an expedited manner are detailed diagrammatically in routes 951 or 981, shown in Figures 62 and 63, respectively and discussed in detail below. If, however, a longer clotting time, that is, in a range of two to eight minutes, is desirous the method to obtain the platelet gel composition ofthe present invention can proceed along the routes 971 and 987, which are also detailed diagrammatically in Figures 63 and 63, respectively and discussed in detail below.
• Phase-one according to the preferred embodiment begins by restoring the clot-forming process.
• an agent capable of reversing the effects of the anticoagulation agent is added back into the first portion ofthe platelet rich plasma 260 stored in vessel 952.
• the restoration agent can be vessel 952 itself or the restoration agent is contained within vessel 952 prior to the introduction of platelet rich plasma 260; however, the restoration agent may also be introduced later.
• the contact activator be a material such as but not limited to glass wool 953 or silica, aluminum, diatomaceous earth, kaolin, etc., or non-wettable surfaces such as plastic, siliconized glass, etc.
• Chemical activators such as kaolin, can also be used to speed up the clotting time; however, their subsequent removal would also be necessary.
• a plastic syringe is the prefe ⁇ ed container used to collect the desired fraction.
• the reversal of the anticoagulant is accomplished using calcium chloride.
• any substance which is known or found to be functionally equivalent to calcium chloride such as, calcium gluconate or calcium carbonate, in restoring the coagulation activity of citrated blood may be used in the practice ofthe present invention.
• calcium chloride is the presently preferred calcium salt for use in the invention, any calcium salt which functions in a similar manner to calcium chloride may be used in the invention.
• any substance which is known or subsequently found to be functionally equivalent to calcium in facilitating these coagulation reactions may be used, either individually or in combination with calcium, in the practice ofthe present invention.
• the anticoagulation agent used was heparin, then heparinase or any other suitable anticoagulant reversing compound would be used to reverse the effect ofthe anticoagulation agent.
• the concentration of the restoration agent used to reverse the anticoagulation will depend in part, upon the concentration of the anticoagulation agent in the platelet rich plasma 260 and the stoichiometry of the chelating and coagulation reactions. However, the concentration of the restoration agent used to reverse the anticoagulation must be sufficient to achieve clot formation.
• a clot 954 Upon restoration of the platelet rich plasma 260 as shown in Figures 62, a clot 954 will naturally form.
• the resulting clot 954 is then triturated by squeezing the clot 954 through glass wool 953 which serves not only as a contact activator but also as a filter, thus expressing thrombin 955.
• a filter 958 having a large micron pore size thereby allowing the removal of clot debris and any activator or solids that are present.
• Filter 958 is positioned at the outlet 956 of vessel 952.
• the thrombin 955 is then mixed with the second portion of platelet rich plasma (PRP) 260 contained within vessel 960 to form the platelet gel composition 970 of the present invention in less than three minutes and in quantities sufficient for clinical use.
• PRP platelet rich plasma
• thrombin formed by the intrinsic pathway or the extrinsic pathway.
• an activating agent such as glass wool
• the yield of autologous thrombin by this method may be low due to incomplete conversion of prothrombin and the inactivation of generated thrombin by fibrin and antithrombin III.
• the addition of modifying agents, such as epsilon aminocaproic acid, to the plasma may improve the yield by reducing the amount of thrombin neutralization.
• thromboplastic material upon which the necessary clotting factors will assemble to maximize the rate of prothrombin conversion.
• the activated platelet membrane provides such a stimulant surface and also enriches the necessary factor V activity by secreting additional factor V during platelet degranulation.
• exogenous lipoprotein and/or thromboplastic material to the plasma environment may also serve to maximize the thrombin generation by activation of both intrinsic and extrinsic clotting cascades. Additional amplification of autologous thrombin generation may also be attained by pretreatment of PRP and/or PPP to block or remove both antithrombin-III and fibrinogen prior to conversion of prothrombin to thrombin.
• thrombin 950 is mixed with the platelet poor plasma 262 of phase-two thereby forming the autologous platelet gel composition 972 of the present invention in less than three minutes.
• a third embodiment ofthe present invention, route 971, shown in Figure 62, contemplates collecting the original quantity of platelet rich plasma (PRP) 260 derived from the anticoagulated whole blood 396 in a container, having a wettable surface, such as glass.
• PRP platelet rich plasma
• the platelet rich plasma 260 is then recalcified and the platelet gel composition 974 forms.
• the desired platelet gel composition 974 will require approximately two to eight minutes to form as opposed to less than a three minute formation as was described in the prefe ⁇ ed embodiment.
• the platelet poor plasma 262 rather then the platelet rich plasma 260, is divided into two portions, as discussed previously in the prefe ⁇ ed embodiment.
• the first portion, used in phase-one, which is approximately l A to Vz the original volume is stored in a vessel 952 having a wettable surface, then the restoration agent, preferably calcium chloride, is added directly to the platelet poor plasma 262.
• the restoration agent preferably calcium chloride
• Surface activation ofthe restored platelet poor plasma 262 occurs as result ofthe vessel's surface and/or the glass wool 953 or other surface or chemical activators and a clot 962 thus forms.
• the resulting clot 962 is triturated, as described previously, and the thrombin 955 is collected.
• Thrombin 955 is then mixed with the platelet rich plasma 260 of phase-two thereby forming the platelet gel sealant composition 973.
• thrombin 955 is mixed with the platelet poor plasma 262 of phase-two thereby forming the platelet gel composition 975 in less than three minutes.
• the sixth embodiment follows route 987, shown in Figure 63 wherein the original quantity of platelet poor plasma 262 is collected in a container having a wettable surface, such as glass. The platelet poor plasma 262 is then recalcified and the platelet gel composition forms.
• the tensile strength ofthe platelet gel compositions of the present invention can be effected by the addition of calcium ions. Consequently, if a stronger bioadhesive sealant composition is desired using the methods discussed above and disclosed in routes 951 and
• the time period necessary for the formation of the platelet gel composition of the present invention is dependent on the quantity of serum added.
• a 1 :4, 1 :2 and 3:4 ratio of serum to platelet rich plasma or platelet poor plasma results in the formation ofthe platelet gel composition in approximately 90, 55 and 30 seconds, respectively.
• thrombin is autocatalytic, it is important that the serum be used within five hours of preparation, preferably within two hours and ideally immediately. Alternatively, the serum can be chilled or frozen indefinitely.
• the platelet gel compositions of this invention may be used for sealing a surgical wound by applying to the wound a suitable amount platelet rich plasma or platelet poor plasma. Moreover, due to the fact that the platelet gel compositions ofthe present invention have been prepared solely from blood components derived from the patient that is to receive the platelet gel there is a zero probability of introducing a new blood transmitted disease to the patient.
• the methods ofthe present invention may be further modified so that the formed platelet gel composition functions not only as a haemostatic agent, but also as an adjunct to wound healing and as a matrix for delivery of drugs and proteins with other biologic activities. For example, it is well known that fibrin glue has a great affinity to bind bone fragments which is useful in bone reconstruction, as in plastic surgery or the repair of major bone breaks.
• autologous bone from a patient can be ground or made into powder or the like, and mixed into the platelet rich plasma obtained in phase-two of the methods of the present invention.
• Autologous thrombin is then mixed in with the platelet rich plasma and bone fragments in an amount sufficient to allow the resulting gel to be applied to the desired locale where it congeals.
• Other materials that may be utilized are, but not limited to, gelatin collagen, degradable polymers, hyaluronic acid, carbohydrates and starches.
• the method ofthe present invention may be modified as follows.
• a wide variety of dmgs and proteins with other biologic activities may be added to the platelet rich plasma of phase-two.
• agents to be added to the platelet rich plasma prior to the addition of the serum include, but are not limited to, analgesic compounds, such as Lidocaine, antibacterial compounds, including bactericidal and bacteriostatic compounds, antibiotics (e.g., adriamycin, erythromycin, gentimycin, penicillin, tobramycin), antifungal compounds, anti-inflammatories, antiparasitic compounds, antiviral compounds, anticancer compounds, such as paclitaxol enzymes, enzyme inhibitors, glycoproteins, growth factors (e.g.
• lymphokines cytokines
• hormones steroids, glucocorticosteroids, immunomodulators, immunoglobulins, minerals, neuroleptics, proteins, peptides, lipoproteins, tumoricidal compounds, tumorstatic compounds, toxins and vitamins (e.g., Vitamin A, Vitamin E, Vitamin B, Vitamin C, Vitamin D, or derivatives thereof). It is also envisioned that selected fragments, portions, derivatives, or analogues of some or all of the above may be used.
• Some of the more successful techniques of evaluating blood clotting and coagulation are the plunger techniques illusfrated by U.S. Pat. Nos. 4,599,219 to Cooper et al, 4,752,449 to Jackson et al, and 5,174,961 to Smith, all of which are assigned to the assignee ofthe present invention, and all of which are inco ⁇ orated herein by reference.
• Automated apparatuses employing the plunger technique for measuring and detecting coagulation and coagulation-related activities generally comprise a plunger sensor cartridge or cartridges and a microprocessor controlled apparatus into which the cartridge is inserted.
• the apparatus acts upon the cartridge and the blood sample placed therein to induce and detect the coagulation-related event.
• the cartridge includes a plurality of test cells, each of which is defined by a tube-like member having an upper reaction chamber where a plunger assembly is located and where the analytical test is ca ⁇ ied out, and a reagent chamber which contains a reagent or reagents.
• the reagents include an activation reagent to activate coagulation of the blood.
• a plug member seals the bottom of a reagent chamber.
• the contents of the reagent chamber are forced into the reaction chamber to be mixed with the sample of fluid, usually human blood or its components.
• An actuator which is a part ofthe apparatus, lifts the plunger assembly and lowers it, thereby reciprocating the plunger assembly through the pool of fluid in the reaction chamber.
• the plunger assembly descends by the force of gravity, resisted by a property of the fluid in the reaction chamber, such as its viscosity.
• the property ofthe sample changes in a predetermined manner as a result ofthe onset or occu ⁇ ence of a coagulation-related activity, the descent rate of the plunger assembly there through is changed. Upon a sufficient change in the descent rate, the coagulation-related activity is detected and indicated by the apparatus.
• Figure 64 the amount of serum, the type of plasma from which the serum was derived, and the type of plasma the serum was mixed with were tested to determine the shortest clotting times.
• the results of which are represented in Figures 66 and 67 the relationship between actual gel time for the platelet gel ofthe present was compared to the clotting time in the cartridge, wherein there is a 0, 30, or 60 minute delay of adding the serum from its generation.
• the third set of experiments the results of which are represented in Figures 68 and 69, studied the effect of calcium addition on actual gel time versus clotting time in the cartridge.
• cc's of platelet rich plasma are drawn into receiving chamber 961 and 3 cc's per PRP or PPP are drawn into receiving chamber 957 which further contains 0.33 cc's of 10% calcium chloride and glass wool. Clotting ofthe contents will occur in two to eight minutes in receiving chamber 957. The clot is then squeezed through optional filter 958 and the semm, produced therefrom, is added to the platelet rich plasma contained in receiving chamber 961 by either mixing or spraying the two components. The platelet rich plasma and the semm will gel within approximately three minutes.
• the inactive blood component and thrombin can be mixed and/or injected into a mold having a desired geometric shape.
• the mold may be constructed of a material having a wettable surface, such as, but not limited to plastic.
• platelet gel of the present invention may be used to temporarily fill, cavities such as but not limited to holes left in the gum from tooth extraction and/or holes left in tissue or bone as a result of injury or surgical procedures.
• the present invention provides a simpler way of introducing platelet gel for specific uses, by providing that the platelet gel be pre-shaped or molded into a beneficial shape prior to being inserted into a cavity.
• the platelet gel may be shaped so as to achieve a basic conical shape. Other shapes such as, but not limited to rods, and rectangles are contemplated by this invention.
• the ability to cause the gel to be more, or less, solid and thus malleable may be achieved during the activation sequence ofthe gel formation.

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• 2002
  • 2002-04-05 AT AT02763954T patent/ATE474668T1/de not_active IP Right Cessation
  • 2002-04-05 DE DE60237091T patent/DE60237091D1/de not_active Expired - Lifetime
  • 2002-04-05 EP EP10007534.0A patent/EP2266705A3/de not_active Withdrawn
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